ATM related kinase ATX, nucleic acids encoding same and methods of use

ABSTRACT

The invention provides an isolated nucleic acid molecule having substantially the same nucleotide sequence as SEQ ID NO:1. Also provided is an isolated oligonucleotide having at least 15 contiguous nucleotides of a nucleotide sequence referenced as SEQ ID NO:11. An isolated polypeptide having substantially the same amino acid sequence as SEQ ID NO:2 is further provided as well as an antibody, or antigen binding fragment thereof, which specifically binds to an ATX polypeptide and has an amino acid sequence as referenced in SEQ ID NO:2. A method for identifying an ATX-modulatory compound is additionally provided. The method consists of measuring the level of an ATX polypeptide in the presence of a test compound, wherein a difference in the level of said ATX polypeptide in the presence of said test compound compared to in the absence of said test compound indicating that said test compound is an ATX-modulatory compound, and wherein said ATX-modulatory compound is not caffeine or wortmannin.

[0001] This application is based on, and claims the benefit of, U.S.Provisional Application No. 60/______ (yet to be assigned), filed Jun.6, 2002, which was converted from U.S. Ser. No. 10/165,216, and which isincorporated herein by reference.

[0002] This invention was made with government support under grantnumber CA76193 awarded by the National Institutes of Health. The UnitedStates Government has certain rights in this invention.

BACKGROUND OF THE INVENTION

[0003] This invention relates generally to the fields of molecularbiology and molecular medicine and more specifically to theidentification of phosphoinositide 3-kinase related kinases (PIKKs)involved in cell cycle control and mRNA surveillance pathways.

[0004] The mitotic cell cycle is the process by which a cell creates anexact copy of its chromosomes and then segregates each copy into twocells. The sequence of events of the cell cycle is regulated such thatcell division does not occur until the cell has completed accurate DNAreplication. To ensure that cells pass accurate copies of their genomeson to the next generation, evolution has overlaid the core cell cyclemachinery with a series of surveillance pathways termed cell cyclecheckpoints. The overall function of these checkpoints is to detectdamaged or abnormally structured DNA, and to coordinate cell cycleprogression with DNA repair.

[0005] Members of the phosphoinositide 3-kinase related kinases (PIKK)family of kinases are involved in cell cycle checkpoints and DNA damagerepair. The PIKK family members identified to date express acarboxylterminal domain that displays significant sequence homology tothe catalytic domains of phosphoinositide (PI) 3-kinases. Indeed, many,but not all of the PIKKs have been shown to possess proteinserine-threonine kinase activities (McMahon et al., Cell 94:363-374(1998); Vassilev et al., Cell 2:869-875 (1998); Grant et al., Cell2:863-867 (1998); Hunter, Cell 83:1-4 (1995); Tibbetts and Abraham,Signaling Networks and Cell Cycle: Themolecular Basis of Cancer andOther Diseases pp. 267-301 (2000)). In mammalian cells, three PIKKfamily members, ATM, ATR, and DNA-dependent protein kinase (DNA-PK),serve as proximal signal transducers in cell-cycle checkpoint and DNArepair pathways (Abraham, Genes & Dev. 15:2177-2196 (2001); Durocher andJackson, Curr. Opin. Cell Biol. 13:2225-231 (2001)). The critical rolesof ATM in orchestrating cellular responses to various forms of stressare underscored by the diverse pathologies associated with thehereditary disorder, ataxiatelangiectasia (A-T) (Crawford, SeminarsinPed. Neuro. 5:287-294 (1998); Rotman and Shiloh, Human Mol. Gen.7:1555-1563 (1998); Rotman and Shiloh, Oncogene 18:6135-6144 (1999)).A-T patients lack functional ATM and develop symptoms including extremesensitivity to irradiation, cerebellar degeneration, oculocutaneoustelangiectasias, gonadal deficiencies, immunodeficiencies, and increasedrisk of cancer (Lehman and Carr, Trends in Genet. 11:375-377 (1995)).Fibroblasts derived from these patients show defects in cell cyclecheckpoints and are defective in their response to irradiation (Painterand Young, Proc. Natl. Acad. Sci. (USA) 77:7315-7317 (1980)).

[0006] In general, the proteins in the PIKK family of kinases playimportant roles in mRNA surveillance and cell cycle progression in orderto insure genetic integrity from generation to generation. Compoundsthat modulate PIKK polypeptides can result in altered progressionthrough the cell cycle leading to increased or decreased cell survival.For example, a PIKK-modulatory compound can make a cell more or lesssusceptible to cell death in the presence of radiation or a cytotoxicagent.

[0007] All cancer cells have a dysfunctional cell cycle and continuethrough the cell cycle in an inappropriate manner, either by failing torespond to negative growth signals or by failing to die in response tothe appropriate signal. In addition, most cancer cells lack genomicintegrity and often have an increased chromosome count compared tonormal cells. Therefore, compounds that inhibit cell cycle checkpointsor DNA damage repair, in combination with the cytotoxic agents, cancause cancer cell death by forcing cancer cells to progress through thecell cycle in the presence of DNA damaging agents such that they undergoevents that lead to cell death.

[0008] Thus, there exists a need to identify additional members of thePIKK family of kinases and compounds that modulate these kinases. Thepresent invention satisfies this need and provides related advantages aswell.

SUMMARY OF THE INVENTION

[0009] The invention provides an isolated nucleic acid molecule havingsubstantially the same nucleotide sequence as SEQ ID NO:1. Also providedis an isolated oligonucleotide having at least 15 contiguous nucleotidesof a nucleotide sequence referenced as SEQ ID NO:11. An isolatedpolypeptide having substantially the same amino acid sequence as SEQ IDNO:2 is further provided as well as an antibody, or antigen bindingfragment thereof, which specifically binds to an ATX polypeptide and hasan amino acid sequence as referenced in SEQ ID NO:2. A method foridentifying an ATX-modulatory compound is additionally provided. Themethod consists of measuring the level of an ATX polypeptide in thepresence of a test compound, wherein a difference in the level of saidATX polypeptide in the presence of said test compound compared to in theabsence of said test compound indicating that said test compound is anATX-modulatory compound, and wherein said ATX-modulatory compound is notcaffeine or wortmannin.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010]FIG. 1A shows the genomic structure of the ATX locus along withclones isolated to date. The black diamonds denote the locations oftranslational stop codons and black bars indicate open reading framesthat give rise to various ATX polypeptides. The lines and symbols belowexon 15 indicate an allelic variant that contains a 27 bp insertionhaving two in-frame stop codons. FIG. 1B shows the location ofN-terminal homology 1 (NH1), NH2, PI3-K catalytic (PI3-Kc),PKC-λ/ι-interacting protein (LIP), and FAT-C (C) domains. The numbersshown indicate % identity/similarity and shading highlights amino acididentity with ATX. A sequence alignment of the PI 3-Kc domains of ATX,CeSMG-1, mTOR, and ATM is shown. FIG. 1C shows immune complex kinaseassays with GST-p53₁₋₇₀, GST-p53₁₋₇₀(S15A), or with GSThUpf11019-1118 assubstrates. The reaction products were immunoblotted with α-HA (lowerpanel). FIG. 1D shows immune complex assays with cells or α-HA-ATXimmunoprecipitates treated with wortmannin.

[0011]FIG. 2A shows clonogenic survival assays after UV exposure. FIG.2B shows clonogenic survival assays after IR exposure. The upper paneldisplays colony outgrowth results from cells transfected with theindicated plasmids, and not exposed to IR. FIG. 2C shows clonogenicsurvival assays of cells treated with ATX-directed antisenseoligonucleotides (AS). The right panel displays colony survival resultsfrom non-irradiated cells treated with the indicated oligonucleotides.

[0012]FIG. 3A shows whole cell extracts resolved by SDS-PAGE andsequentially immunblotted with the indicated antibodies. FIG. 3B showsextracts of transfected cells separated by SDS-PAGE and sequentiallyimmunoblotted with the indicated antibodies. The p53 phosphoserine-15specific antibody is designated α-pSer15. FIG. 3C shows extracts ofcells treated with S or AS oligonucleotides and analyzed as described inpanel A. FIG. 3D shows cell cycle progression in AS-transfected cellsexamined by flow cytometry. The table shows the percentages of cells ineach cell cycle phase. The right panel shows immunoblot analyses fromthe same cell population. FIG. 3E shows an effect of caffeine onAS-induced cell cycle defects. The table shows percentages of cells ineach cell-cycle phase, plus the ratio of G2/M to G1 cells for eachsample. The right panel shows immunoblotting results from the same cellpopulations.

[0013]FIG. 4A shows an effect of ATX overexpression on basal viability.Cell densities of the scanned images from each sample were obtained withthe ImagePro Plus software program. FIG. 4B shows an effect of ATXoverexpression on radiosensitivity. Surviving cells were quantitated asdescribed in panel except that arbitrary unit values for each group werenormalized to the corresponding nonirradiated control.

[0014]FIG. 5A shows cells transfected with GAL4 or GAL4-hUpf11019-1118expression constructs with the indicated samples treated withwortmannin. The right panel shows phosphatase treatment. The solubleproteins were separated by SDS-PAGE and immunoblotted with α-GAL4 mAb.The arrow indicates the uppermost band of the phosphorylatedGAL4-hUpf11019-1118 reporter protein. FIG. 5B shows an effect of ATMKIor ATXKI expression on UV stimulation of GAL4-hUpf1 phosphorylation.

[0015]FIG. 6A shows an effect of HA-ATXKI or HAATMKI expression on NMD.Nuclear RNA was isolated from transfected cells and β-globin and MUPmRNAs were quantitated by RT-PCR and PhosphorImaging. For each pair oftransfections, the level of Globin mRNA was normalized to the level ofMUP mRNA and expressed below each lane as a percentage of the normalizedlevel of Globin Norm mRNA, which was defined as 100. FIG. 6B shows aneffect of ATX AS oligonucleotide on NMD.

[0016]FIG. 7 shows the effect of small-interfering (si) RNA reduction ofATX on response to ionizing radiation (IR). U20S cells transfected withsi RNA directed against luciferase (control) or ATX were treated withdifferent doses of ionizing radiation and lysed after 1 hour or 18hours. Cellular extracts were separated by SDS-PAGE and immunoblottedwith an anti-phospho-Histone H2AX antibody. The level of a controlpolypeptide, α-tubulin, is shown in the lower panel for comparison.

DETAILED DESCRIPTION OF THE INVENTION

[0017] This invention is directed to isolated ATX nucleic acids andpolypeptides. ATX is a novel PIKK kinase family member that participatesin stress-induced p53 and cell cycle checkpoint activation in cellsexposed to DNA damaging agents. In addition, ATX can activate theintrinsic non-sense mediated mRNA decay (NMD) pathway in these cells.The invention is also directed to methods of identifying ATX-modulatorycompounds and using these compounds to modulate cell survival. Compoundsthat modulate cellular survival can be useful in the treatment ofdiseases characterized by excessive cell growth or excessive cell death.

[0018] In one embodiment, an expressed sequence tag (EST) with homologyto a conserved region in the catalytic domains of PIKK family memberswas used to isolate a full-length cDNA encoding a novel member of thePIKK family, termed ATX. The ATX polypeptide was detected in both thenucleus and cytoplasm of human cells, and formed nuclear foci uponexposure to UV light. In addition, the cell cycle regulatory proteinsp53 and hUpf1 were found to be phosphorylated by ATX. Furthermore, thereduction of endogenous ATX in a cell using anti-sense oligonucleotidesresulted in decreased survival of cells, and decreased phosphorylationand stabilization of p53 in cells exposed to UV light. Similar to otherPIKK family members, ATX activity was inhibited by wortmannin andcaffeine.

[0019] As used herein, the term “ATX polypeptide” refers to apolypeptide with substantially the same amino acid sequence as thatshown in SEQ ID NO:2 (human ATX). “Substantially the same amino acidsequence” is intended to mean an amino acid sequence contains aconsiderable degree of sequence identity or similarity, such as at least70%, 80%, 90%, 95%, 98%, or 100% sequence identity or similarity, to areference amino acid sequence. Substantially the same amino acidsequence includes conservative and non-conservative amino acid changes,gaps, and insertions to an amino acid sequence. Conservative andnon-conservative amino acid changes, gaps, and insertions to an aminoacid sequence can be compared to a reference sequence using availablealgorithms and programs such as the Smith-Waterman algorithm and theBLAST homology search program (Altschul et al., J. Mol. Biol.215:403-410 (1990)).

[0020] It is understood that a fragment of ATX can be sufficient inorder to produce an ATX activity. Activities associated with ATXinclude, for example, kinase activity, cell cycle checkpoint activity,and NMD activity. For example, fragments of ATX which retainsubstantially an activity of the entire polypeptide are included withinthe definition. Fragments can include, for example, amino terminal,carboxyl terminal, or internal deletions of a full length ATXpolypeptide. In addition, fragments can include domains of a full lengthATX polypeptide, such as for example, a kinase domain, NH1 domain, NH2domain, or LIP domain. A fragment can contain, for example, at leastabout 10, 100, 500, 1,000, 1,500, 2,000, 2,500, 3,000, 3,500 or morecontiguous or non-contiguous amino acid residues of a full-length ATXpolypeptide. ATX polypeptide fragments include the fragments describedabove, but excludes fragments KIAA0421 (Accession number AB007881),KIAA0220 (Accession number D86974), and LIP (Accession number U32581),which are present in databases. Polypeptide fragments can be generatedusing a variety of methods. For example, polypeptide fragments can begenerated using recombinant DNA methods, enzymatic cleavage, or chemicalcleavage of larger polypeptides.

[0021] It is understood that limited modifications to the ATXpolypeptide can be made without destroying an activity of ATX. Forexample, ATX is intended to include other ATX family members such asthose polypeptides that are found to exhibit the above sequencehomologies. Such members include, for example, homologs of ATX that canbe cloned from other organisms such as monkeys, cows, rats, mice,chickens, frogs, flies or worms. The sequence of possible homologs ofhuman ATX are available in nucleotide databases.

[0022] Various modifications of the ATX primary amino acid sequence canresult in polypeptides having substantially equivalent, decreased, orenhanced function as compared to the sequence set forth as SEQ ID NO:2.Those skilled in the art recognize that such modifications can bedesirable at times in order to enhance the bioactivity, bioavailabilityor stability of ATX, or to facilitate its synthesis or purification.Contemplated amino acid substitutions to the native sequence of ATX caninclude, for example, conservative changes, wherein a substituted aminoacid has similar structural or chemical properties such as replacementof a polar amino acid with another polar amino acid or replacement of acharged amino acid with a similarly charged amino acid. Those skilled inthe art also recognize that nonconservative changes such as replacementof an uncharged polar amino acid with an non-polar amino acid orreplacement of a charged amino acid with an uncharged polar amino acid,can also be made without affecting a function of ATX. In addition, avariety of polypeptide modifications are known in the art forconstraining the structure of polypeptides to enhance stability orbinding (Cabezas and Satterthwait, J. Am. Chem. Soc. 121:3862-3875(1999); Stanfield et al., Structure 7:131-142 (1999)).

[0023] A polypeptide can be modified by naturally occurringmodifications such as post-translational modifications, includingphosphorylation, lipidation, prenylation, sulfation, hydroxylation,acetylation, addition of carbohydrate, addition of prosthetic groups orcofactors, formation of disulfide bonds, proteolysis, assembly intomacromolecular complexes, and the like. Chemical modifications of thepolypeptide such as, for example, alkylation, acylation, carbamylation,and iodination can also be used to modify an ATX polypeptide. Inaddition, various molecules, such as other polypeptides, carbohydrates,or lipids, or small molecules can be attached to ATX including fragmentsof ATX. For example, ATX can contain a label moiety, a sequence such asa FLAG epitope, or be fused to another polypeptide such as a DNA bindingdomain.

[0024] Those skilled in the art can determine which residues and whichregions of a ATX sequence are likely to be tolerant of modification andstill retain an activity associated with ATX. For example, amino acidsubstitutions or chemical or enzymatic modifications at residues thatare less well conserved between species are more likely to be toleratedthan substitutions at highly conserved residues. Accordingly, analignment can be performed among ATX sequences of various species todetermine residues and regions in which modifications are likely to betolerated (FIG. 1B). Additional guidance for determining residues andregions of ATX likely to be tolerant of modification is provided bystudies of ATX fragments and variants. In addition, it can be useful tomodify ATX in a way that destroys an activity associated with ATX. Forexample, as disclosed herein, the mutation of an aspartic acid to analanine at conserved residue Asp-2195 in the ATX kinase domain generatesa kinase-inactive version of ATX.

[0025] As used herein, the term “level” in reference to a level of anATX nucleic acid or polypeptide refers to the amount, accumulation, orrate of synthesis of a molecule or to the amount or rate of an activityassociated with the molecule. A level can be represented, for example,by the amount or synthesis rate of messenger RNA (mRNA) encoded by agene, the amount or synthesis rate of polypeptide corresponding to agiven amino acid sequence encoded by a gene, or the amount or synthesisrate of a biochemical form of a molecule accumulated in a cell,including, for example, the amount of particular post-syntheticmodifications of a molecule such as a polypeptide or nucleic acid. Inaddition, a level can be represented, for example, by the extent ofphosphorylation of a substrate molecule or by the amount of an activitysuch as cell cycle checkpoint activity, NMD activity or ability toinduce cell death or cell survival. The term can be used to refer to anabsolute amount of a molecule or activity in a sample or to a relativeamount of the molecule or activity, including amounts and activitiesdetermined under steady-state or non-steady-state conditions. Forexample, the expression level of a molecule can be determined relativeto a control component molecule in a sample.

[0026] As used herein, the term “p53” is intended to mean a polypeptidewith substantially the same amino acid sequence as that shown in SEQ IDNO:4 (human p53). As described above for ATX, it is understood that p53includes fragments of the full length p53 polypeptide. For example, theamino terminal 70 amino acids of p53 (p53 1-70) can be used in themethods of the invention as a substrate for ATX kinase activity. Also,for example, a fragment of p53 that includes the LSQE sequence locatedat amino acids 14 to 17 of p53 can be used as a substrate for ATX kinaseactivity. In addition, as described above for ATX, a p53 polypeptideincludes p53 from species other than humans, and includes modificationsto the p53 polypeptide including conservative and non-conservative aminoacid changes, post-translational modifications and chemicalmodification. Also, as described for ATX, a p53 polypeptide can containadditional sequences such as a known epitope or a label moiety.

[0027] The term “specifically binds” is intended to mean the moleculewill have an affinity for the target molecule that is measurably higherthan its affinity for a non-specific interaction. For example, a nucleicacid can specifically bind to another nucleic acid by complementary basepairing between the nucleotides. In addition, a polypeptide such as anantibody that specifically binds another polypeptide will have anaffinity for the target polypeptide or antigen that is measurably higherthan its affinity for a non-specific interaction. Furthermore, acompound such as a small organic molecule can specifically bind to atarget molecule with an affinity that is measurably higher than itsaffinity for a non-specific interaction. Binding affinity can be low orhigh affinity so long as the binding is sufficient to be detectable. Forexample, a compound can bind ATX with a binding affinity (Kd) of about10⁻⁴ M or less, 10⁻⁵ M or less, 10⁻⁶ M or less, about 10⁻⁷ M or less,including about 10⁻⁸ M or less, such as 10⁻⁹ M or less. Several methodsfor detecting or measuring nucleotide, polypeptide, and other compoundbinding are well known in the art and disclosed herein.

[0028] As used herein, the term “compound” is intended to mean anisolated macromolecule of natural or synthetic origin that can beassayed using the methods of the invention. A compound includes, forexample, a polypeptide, peptidomimetic, non-peptidyl compound,carbohydrate, lipid, an antibody or antibody fragment, a small organicor inorganic molecule, or a nucleotide sequence including an aptamer,antisense oligonucleotide, interfering RNA or ribozyme. For example, acompound can be an isolated cDNA sequence. A compound can have a knownor unknown structure. A compound can be isolated or be part of apopulation of compounds such as a library. For example, a compound canbe a small organic compound obtained from a combinatorial chemicallibrary. A library of compounds can be a random collection of compoundsor can be rationally designed based on a physical characteristic. Acompound which is assayed in the methods of the invention can be calleda “test compound” and if the test compound has the ability to modulatethe level of ATX it can be called an “ATX-modulatory compound.” Onecompound or more than one compound can be used in the methods of theinvention.

[0029] As used herein, a “stressor agent” is any agent that can induce astress response pathway within a cell. Several stressor agents are knownin the art such as UV light, ionizing radiation, reactive oxygenintermediates, cytotoxic agents, and replicational stress imposed by DNAreplication inhibitors including, for example, hydroxyurea andaphidicolin. In addition, environmental conditions such as excessiveheat can induce a stress-response pathway within a cell resulting in,for example, the induction of heat shock proteins. Stress responsepathways include DNA repair pathways, non-sense mediated mRNA decay(NMD), heat shock pathways, the induction of apoptosis, activation ofthe NFkB transcription factor, activation of the stress-activated MAPkinase pathways including, for example, JNK and p38 pathways, andactivation of ubiquitin-dependent proteolysis.

[0030] As used herein, the term “non-sense mediated messenger RNA (mRNA)decay (NMD)” is intended to mean the surveillance mechanism within cellswhereby imperfect mRNAs that contain premature translation terminationcodons are preferentially degraded. These imperfect mRNAs can result inpolypeptides that are nonfunctional or have altered function such asgain-of function or dominant negative mutations.

[0031] As used herein, the term an “amount effective” or “effectiveamount” when used in reference to a compound that modulates cellsurvival or growth is intended to mean an amount of the compound ormolecule sufficient to increase or decrease cell survival or growth.Modulation also includes induction of cell survival or growth orcomplete blockage of cell survival or growth. In addition, an effectiveamount of a compound is intended to mean an amount of the compound thatis sufficient to treat or reduce the severity of a condition in anaffected subject.

[0032] The invention provides an isolated nucleic acid molecule havingsubstantially the same nucleotide sequence as SEQ ID NO:1. In addition,the invention provides an isolated nucleic acid molecule havingsubstantially the same nucleotide sequence as SEQ ID NO:1 where thenucleic acid molecule encodes an ATX polypeptide containing an aminoacid sequence shown in SEQ ID NO:2. For example, the invention providesan isolated nucleic acid molecule containing the sequence shown in SEQID NO:1.

[0033] Substantially the same nucleic acid sequence is intended to meana nucleic acid sequence contains a considerable degree of sequenceidentity or similarity, such as at least 70%, 80%, 90%, 95%, 98%, or100% sequence identity or similarity, to a reference nucleic acidsequence. Substantially the same nucleic acid sequence includes nucleicacid changes, gaps, and insertions to an nucleic sequence. Nucleic acidchanges, gaps, and insertions to a nucleic acid sequence can be comparedto a reference sequence using available algorithms and programs such asthe Smith-Waterman algorithm and the BLAST homology search program(Altschul et al., J. Mol. Biol. 215:403-410 (1990)).

[0034] Isolated nucleic acid molecules include DNA sequences and RNAtranscripts, both sense and complementary anti-sense strands, includingsplice variants thereof encoding ATX polypeptides. An isolated nucleicacid molecule can contain a double stranded molecules or single strandedmolecules, including RNA as well as coding and noncoding DNA. DNAsequences of the invention include genomic and cDNA sequences as well aswholly or partially chemically synthesized DNA sequences. Genomic DNA ofthe invention comprises the protein coding region for a polypeptide ofthe invention and includes allelic variants of the preferred nucleicacid of the invention. Genomic DNA of the invention is distinguishablefrom genomic DNAs encoding polypeptides other than ATX in that itincludes an ATX protein coding region found in ATX-encoding cDNA of theinvention. Genomic DNA of the invention can be transcribed into RNA, andthe resulting RNA transcript can undergo one or more splicing eventswherein one or more introns of the transcript are removed, or “splicedout.” Peptide nucleic acids (PNAs) encoding a polypeptide of theinvention are also contemplated (Corey, TIBTech 15:224-229 (1997)). PNAsare DNA analogs containing neutral amide backbone linkages that areresistant to DNA degradation enzymes and which bind to complementarysequences at higher affinity than analogous DNA sequences as a result ofthe neutral charge on the backbone of the molecule.

[0035] RNA transcripts that can be spliced by alternative mechanisms,and therefore be subject to removal of different RNA sequences but stillencode an ATX polypeptide, are referred to in the art as splice variantswhich are embraced by the invention. Splice variants comprehended by theinvention therefore are encoded by the same DNA sequences but arise fromdistinct mRNA transcripts. Allelic variants are known in the art to bemodified forms of a wild type gene sequence, the modification resultingfrom recombination during chromosomal segregation or exposure toconditions which give rise to genetic mutation. Allelic variants, likewild type genes, are inherently naturally occurring sequences (asopposed to non-naturally occurring variants which arise from in vitromanipulation).

[0036] An allelic variant of ATX is disclosed herein as SEQ ID NO:5.This form of ATX is produced as the result of allelic variation in exon15 which leads to the insertion of 27 nucleotides beginning atnucleotide 1427 (FIG. 1A). This sequence alteration causes the insertionof two in-frame stop codons and the use of the next available ATG codonin exon 16 as the translational stat site, resulting in anamino-terminally truncated or short form of ATX. A form of ATX that issimilar to the long form of ATX disclosed herein (SEQ ID NO:1) isreferenced as SEQ ID NO:7. This form of ATX has exon 5 spliced to exon 6which results in a different N-terminus and 8 additional amino acids inthe resulting polypeptide (FIG. 1A). In the experiments disclosed hereinclones that were isolated with exon 5 frequently contained exon 3 whichplace an in-frame stop codon at the 3′ end of this DNA (Example 1). Thelongest form of ATX (SEQ ID NO:9) was isolated, however the exon 3associated stop codon was present in this transcript as well.

[0037] In addition to genomic DNA, isolated nucleic acids include cDNA.cDNA can be obtained through reverse transcription of an RNA nucleicacid encoding ATX, followed by second strand synthesis of acomplementary strand to provide a double stranded DNA. In addition,nucleic acid molecules can be chemically synthesized meaning produced bypurely chemical, as opposed to enzymatic, methods. Wholly chemicallysynthesized DNA sequences are produced entirely by chemical means, andpartially synthesized DNAs are those where only portions of theresulting DNA were produced by chemical means.

[0038] ATX nucleic acid molecules include homologs of the human ATXsequence. Species homologs in general share at least 50%, at least 60%,at least 70%, at least 80%, at least 90%, at least 95% or at least 99%homology with a human DNA of the invention. ATX nucleic acids includespecies homologs of the human ATX sequence, but exclude a mouse EST thatcontains a sequence homologous to the 3′ part of ATX is (GenBankAccession Number BC024431) and a Macaca fascicularis brain cDNA cloneQf1A-15747 (accession number AB056380).

[0039] The invention also provides anti-sense oligonucleotides based onSEQ ID NO:1. For example, the invention provides an isolatedoligonucleotide having at least 15 contiguous nucleotides of thenucleotide sequence 5′-AGCAAGCTCCCTCCTGTCTC-3′ (SEQ ID NO:11). Theoligonucleotide shown in SEQ ID NO:11 is an ATX anti-senseoligonucleotide that has been shown herein to decrease the level of ATXin a cell (Example 5).

[0040] Nucleic acids of the invention also permit identification andisolation of nucleic acid encoding related ATX polypeptides by wellknown techniques including Southern hybridization, Northernhybridization, and polymerase chain reaction (PCR). Examples of relatednucleic acids include human and non-human nucleic acid sequences,including allelic variants, as well as nucleic acids encodingpolypeptides homologous to ATX and structurally related polypeptidessharing one or more biological, immunological, or physical properties ofATX.

[0041] The invention provides a method for detecting an ATX nucleic acidmolecule in a sample, by contacting the sample with an ATX nucleic acidmolecule under conditions that allow specific hybridization to ATXnucleic acid, and detecting the specific hybridization. In addition, theinvention provides a method for detecting an ATX nucleic acid moleculein a sample, by contacting a nucleic acid fraction derived from thesample with a PCR primer pair set under conditions that allowamplification of an ATX nucleic acid, and detecting amplified ATXnucleic acid. Kits for detecting ATX nucleic acids based on thesemethods are provided as well.

[0042] Fragments of ATX nucleic acid molecules are useful in theinvention, for example, as probes for detection of full length or otherfragment ATX nucleic acids. A nucleic acid fragment can include forexample 5′, 3′, or internal deletions of a full length ATX nucleic acidsequence. For example, the invention provides an isolated ATX nucleicacid molecule as referenced in SEQ ID NO:5. Alternatively, the inventionprovides ATX nucleic acid fragments other than the fragment asreferenced in SEQ ID NO:5. For example, the invention provides ATXnucleic acid fragments that contain carboxyl terminal deletions of afull length ATX polypeptide. In addition, fragments can include domainsof a full length ATX nucleic acid sequence, for example, a kinasedomain, NH1 domain, NH2 domain, or LIP domain. A fragment can contain,for example, at least about 10, 100, 1,000, 2,500, 5,000, 7,500, 10,000,12,500 or more contiguous or non-contiguous nucleic acid residues of afull-length ATX nucleic acid sequence. ATX nucleic acid fragmentsinclude the fragments described above, but excludes fragments KIAA0421(Accession number AB007881), KIAA0220 (Accession number D86974), and LIP(Accession number U32581), which are present in databases. One or morefragment nucleic acids can be included in kits that are used to detectthe presence of a nucleic acids encoding ATX, or used to detectvariations in a nucleic acid sequence encoding ATX, includingpolymorphisms, for example, single nucleotide polymorphisms.

[0043] The nucleic acids of the invention can contain heterologoussequences that are not part of the ATX-encoding sequences in nature. Theheterologous nucleic acid sequence can be separated from the ATX-codingsequence by an encoded cleavage site that will permit removal of non-ATXpolypeptide sequences from the expressed fusion protein. Heterologousnucleic acids sequences can include sequences encoding epitopes, such aspoly-histidine sequences, FLAG tags, glutathione-S-transferase,thioredoxin, and maltose binding protein domains, that facilitatepurification of the fusion protein. In addition heterologous nucleicacids can encode domains, such as leucine zipper motifs, that promotemultimer formation between the fusion protein and itself or otherproteins or immunoglobulins or fragments thereof that can enhancecirculatory half-life of the encoded protein.

[0044] The nucleic acid molecules of the invention also include DNAsequences encoding ATX species that hybridize under highly or moderatelystringent conditions to the non-coding strand, or complement, of thenucleic acid in SEQ ID NO: 1. ATX-encoding nucleic acids of theinvention include a) the nucleic acid sequence set out in SEQ ID NO: 1;b) nucleic acids encoding a polypeptide encoded by the nucleic acid of(a), and c) nucleic acids that hybridize to the complement of thenucleic acids of (a) or (b) under moderately or highly stringentconditions. Exemplary high stringency conditions include a final wash in0.2×SSC/0.1% SDS at 65° C. to 75° C., and exemplary moderate stringencyconditions include a final wash at 2× to 3×SSC/0.1% SDS at 65° C. to 75°C. It is understood in the art that conditions of equivalent stringencycan be achieved through variation of temperature and buffer, or saltconcentration as described in Ausubel, et al. (Eds.), Protocols inMolecular Biology, John Wiley & Sons (1994). Modifications inhybridization conditions can be empirically determined or preciselycalculated based on the length and the percentage of guanosine/cytosine(GC) base pairing of the probe.

[0045] The invention also provides a vector containing the isolated ATXnucleic acid molecules described above. For example, the inventionprovides a vector containing an isolated nucleic acid molecule havingsubstantially the same nucleotide sequence as SEQ ID NO:1.

[0046] Vectors include autonomously replicating recombinant expressionconstructs such as plasmid and viral DNA vectors. The invention includesvectors where ATX-encoding nucleic acids are operatively linked to anendogenous or exogenous promoter, enhancer, or operator sequence and atranscription terminator sequence. Promoter and enhancer sequences aregenerally selected for the ability to increase gene expression, whileoperator sequences are generally selected for the ability to regulategene expression. It is understood in the art that the choice of hostcell is relevant to selection of an appropriate regulatory sequence.Vectors used in the invention can also include sequences encoding one ormore selectable markers that permit identification of host cells bearingthe construct. Vectors can also include sequences that facilitatehomologous recombination in a host cell.

[0047] Suitable vectors for expression in prokaryotic or eukaryoticcells are well known to those skilled in the art (see, for example,Ausubel et al., supra, 1999). Vectors useful for expression ineukaryotic cells can include, for example, regulatory elements includingthe SV40 early promoter, the cytomegalovirus (CMV) promoter, the mousemammary tumor virus (MMTV) steroid-inducible promoter, Moloney murineleukemia virus (MMLV) promoter, and the like. A vector can include, forexample, viral vectors such as a bacteriophage, a baculovirus or aretrovirus; cosmids or plasmids; and, particularly for cloning largenucleic acid molecules, bacterial artificial chromosome vectors (BACs)and yeast artificial chromosome vectors (YACs). Such vectors arecommercially available, and their uses are well known in the art. Oneskilled in the art will know or can readily determine an appropriatepromoter for expression in a particular host cell. For example, asdisclosed herein, the long form of ATX can be sub-cloned into pcDNA 3.1with an HA tag and transfected using Fugene 6 into human embryonickidney 293T cells (Example 2 and Example 5).

[0048] Vectors useful for expression of an ATX polypeptide can contain aregulatory element that provides tissue specific or inducible expressionof an operatively linked nucleic acid. Such inducible systems, include,for example, tetracycline inducible system (Gossen & Bizard, Proc. Natl.Acad. Sci. USA, 89:5547-5551 (1992); Gossen et al., Science,268:1766-1769 (1995); Clontech, Palo Alto, CA)); metallothioneinpromoter induced by heavy metals; insect steroid hormone responsive toecdysone or related steroids such as muristerone (No et al., Proc. Natl.Acad. Sci. USA, 93:3346-3351 (1996); Yao et al., Nature, 366:476-479(1993); Invitrogen, Carlsbad, Calif.); mouse mammory tumor virus (MMTV)induced by steroids such as glucocortocoid and estrogen (Lee et al.,Nature, 294:228-232 (1981); and heat shock promoters inducible bytemperature changes.

[0049] In addition, viral vectors such as retroviral, adenovirus,adeno-associated virus, lentivirus, and herpesvirus vectors can be usedto express ATX polypeptides into a cell. Viral based systems provide theadvantage of being able to introduce relatively high levels of aheterologous nucleic acid into a variety of cells. Additionally, suchviruses can introduce heterologous DNA into nondividing cells. Viralvectors include, for example, Herpes simplex virus vectors (U.S. Pat.No. 5,501,979), Vaccinia virus vectors (U.S. Pat. No. 5,506,138),Cytomegalovirus vectors (U.S. Pat. No. 5,561,063), Modified Moloneymurine leukemia virus vectors (U.S. Pat. No. 5,693,508), adenovirusvectors (U.S. Pat. Nos. 5,700,470 and 5,731,172), adeno-associated virusvectors (U.S. Pat. No. 5,604,090), constitutive and regulatableretrovirus vectors (U.S. Pat. Nos. 4,405,712; 4,650,764 and 5,739,018,respectively), papilloma virus vectors (U.S. Pat. Nos. 5,674,703 and5,719,054), and the like.

[0050] The invention further provides a host cell containing anATX-encoding vector as described above. For example, the inventionprovides a host cell that contains a vector which contains an isolatednucleic acid molecule having substantially the same nucleotide sequenceas SEQ ID NO:1. Host cells include prokaryotic and eukaryotic cells.Nucleic acids of the invention can be introduced into the host cell aspart of a circular plasmid, or as linear DNA having an isolated proteincoding region or a viral vector. Methods for introducing DNA into thehost cell are well known in the art and include transformation,transfection, electroporation, nuclear injection, or fusion withcarriers such as liposomes, micelles, ghost cells, protoplasts, andother transformed cells. Detailed procedures for these methods can befound in Sambrook et al., Molecular Cloning: A Laboratory Manual (ColdSpring Harbor Laboratory Press, 1989) and the references cited therein).Expression systems of the invention include bacterial, yeast, fungal,plant, insect, invertebrate, and mammalian cells systems.

[0051] Useful mammalian expression vectors and methods of introducingsuch vectors into mammalian cells either ex vivo or in vivo, forexpression of the encoded polypeptide, are well known in the art. Forexample, a plasmid expression vector can be introduced into a cell bycalcium-phosphate mediated transfection, DEAE-Dextran-mediatedtransfection, lipofection, polybrene- or polylysine-mediatedtransfection, electroporation, or by conjugation to an antibody,gramacidin S, artificial viral envelopes or other intracellularcarriers. A viral expression vector can be introduced into a cell in anexpressible form by infection or transduction, for example, or byencapsulation in a liposome.

[0052] The invention also provides a method of producing an ATXpolypeptide by a) growing the host cell described above under conditionsappropriate for expression of the ATX polypeptide, and b) isolating theATX polypeptide from the host cell or host cell growth medium. Thismethod can be used to produce ATX polypeptide, for example, as a sourceof immunogen for the development of antibodies specifically reactivewith ATX.

[0053] ATX polypeptide isolated from the cells or from the medium inwhich the cells are grown by purification methods known in the art, forexample, conventional chromatographic methods including immunoaffinitychromatography, receptor affinity chromatography, hydrophobicinteraction chromatography, lectin affinity chromatography, sizeexclusion filtration, cation or anion exchange chromatography, highpressure liquid chromatography (HPLC), reverse phase HPLC, and the like.Still other methods of purification include those wherein the desiredprotein is expressed and purified as a fusion protein having a specifictag, label, or chelating moiety that is recognized by a specific bindingpartner or agent. The purified protein can be cleaved to yield thedesired protein, or be left as an intact fusion protein.

[0054] The DNA sequence information provided by the present inventionalso makes possible the development through, for example, homologousrecombination or “knock-out” strategies of animals that fail to expressfunctional ATX or that express a variant of ATX (Capecchi, Science244:1288-1292 (1989)). Such animals are useful as models for studyingthe in vivo activities of ATX and modulators of ATX.

[0055] The invention provides an isolated polypeptide containingsubstantially the same amino acid sequence as SEQ ID NO:2. For example,the invention provides a polypeptide containing an amino acid sequenceas referenced in SEQ ID NO:2. The sequence shown in SEQ ID NO:2corresponds to the “long form” of ATX (FIG. 1A).

[0056] As described further above, an isolated ATX polypeptide includesconservative and non-conservative amino acid changes to the sequenceshown in SEQ ID NO:2. In addition, an isolated ATX polypeptide includesspecies homologs and fragments of ATX. For example, the inventionprovides an isolated ATX polypeptide fragment as referenced in SEQ IDNO:6. Alternatively, the invention provides ATX polypeptide fragmentsother than the fragment as referenced in SEQ ID NO:6. For example, theinvention provides ATX polypeptide fragments that contain carboxylterminal deletions of a full length ATX polypeptide. Furthermore, an ATXpolypeptide can contain polypeptide modifications or heterologoussequences such as an epitope tag. Polypeptides of the invention can beisolated from natural cell sources, chemically synthesized, or producedby recombinant procedures involving the host cells of the invention.

[0057] The invention provides an antibody, or antigen binding fragmentthereof, which specifically binds to an ATX polypeptide containing anamino acid sequence as referenced in SEQ ID NO:2. Antibodies include,for example, monoclonal and polyclonal antibodies, single chainantibodies, chimeric antibodies, bifunctional or bispecific antibodies,humanized antibodies, human antibodies, and complementary determiningregion (CDR)-grafted antibodies, including compounds which include CDRor antigen-binding sequences, which specifically bind to a polypeptideof the invention. Antibody fragments, including Fab, Fab′, F(ab′)₂, andFv, are also provided by the invention. Screening assays to determinebinding specificity or exclusivity of an antibody of the invention arewell known in the art (see Harlow et al. (Eds), Antibodies A LaboratoryManual; Cold Spring Harbor Laboratory; Cold Spring Harbor, N.Y. (1988)).

[0058] Antibodies that recognize and bind fragments of the ATXpolypeptides of the invention are also contemplated, provided that theantibodies specifically bind ATX polypeptides. As with antibodies thatare specific for full length ATX polypeptides, antibodies of theinvention that recognize ATX fragments are those which can distinguishATX polypeptides from other PIKK polypeptides despite inherent sequenceidentity, homology, or similarity found in the family of proteins.

[0059] Antibodies of the invention can be produced using any method wellknown in the art, using any polypeptide, or immunogenic fragmentthereof, of the invention. Immunogenic polypeptides can be isolated fromnatural sources, from recombinant host cells, or can be chemicallysynthesized. For example, as disclosed herein, antibodies specificallyreactive with ATX were generated using glutathione S-transferase (GST)fusion proteins containing ATX amino acids 2281-2339 (anti-ATX-Ab-1) oramino acids 1691-1790 (anti-ATX-Ab-2) (Example 2). Polypeptide of theinvention can also be conjugated to a hapten such as keyhole limpethemocyanin (KLH) in order to increase immunogenicity. Methods forsynthesizing such peptides are known in the art, for example, as in R.P. Merrifield, J. Amer. Chem. Soc. 85: 2149-2154 (1963); J. L.Krstenansky, et al., FEBS Lett. 211:10 (1987). Antibodies to apolypeptide of the invention can also be prepared through immunizationusing a nucleic acid of the invention, as described in Fan et al., Nat.Biotech. 17:870-872 (1999). DNA encoding a polypeptide can be used togenerate antibodies against the encoded polypetide following topicaladministration of naked plasmid DNA or following injection, for example,intramuscular injection, of the DNA.

[0060] Non-human antibodies can be humanized by any methods known in theart. In one method, the non-human CDRs are inserted into a humanantibody or consensus antibody framework sequence. Further changes canthen be introduced into the antibody framework to modulate affinity orimmunogenicity. Antibodies of the invention further include plasticantibodies or molecularly imprinted polymers (MIPs) (Haupt and Mosbauch,TIBTech 16:468-475 (1998)). Antibodies of this type can be useful inimmunoaffinity separation, chromatography, solid phase extraction,immunoassays, for use as immunosensors, and for screening chemical orbiological libraries. Advantages of antibodies of this type are that noanimal immunization is required, the antibodies are relativelyinexpensive to produce, they are resistant to organic solvents, and theyare reusable over long period of time.

[0061] The invention provides a method for detecting ATX polypeptide ina sample by contacting the sample with an ATX antibody under conditionsthat allow specific binding of the antibody to the polypeptide anddetecting the bound antibody. Antibodies of the invention can alsoinclude one or more labels that permit detection of the antibody andantibody binding. Labels can include, for example, radioactivity,fluorescence (or chemiluminescence), one of a high affinity binding pair(such as biotin/avidin), enzymes, or combinations of one or more ofthese labels. Antibodies of the invention are also useful, for example,for therapeutic purposes (by modulating activity of ATX), diagnosticpurposes to detect or quantitate ATX, as well as purification of ATX.Kits containing an antibody or antibodies of the invention are alsoprovided.

[0062] The DNA and amino acid sequence information provided by thepresent invention also makes possible the systematic analysis of thestructure and function of ATX. DNA and amino acid sequence informationfor ATX also permits identification of compounds with which an ATXpolypeptide or nucleic acid will interact. Methods to identify compoundsthat bind to ATX include solution assays, in vitro assays where ATXpolypeptides are immobilized, and cell based assays. Identification ofcompounds that bind ATX polypeptides provides potential targets fortherapeutic or prophylactic intervention in pathologies associated withATX biological activity.

[0063] The invention provides a method for identifying a compound thatspecifically binds to an ATX polypeptide of the invention, by a)contacting the ATX polypeptide with a compound, and b) determiningspecific binding of the compound to said ATX polypeptide. As describedfurther above, the term compound includes macromolecules of natural orsynthetic origin including, for example, a polypeptide, peptidomimetic,non-peptidyl compound, carbohydrate, lipid, and antibody or antibodyfragment, a small organic or inorganic molecule, or a nucleic acidincluding an aptamer.

[0064] Identification of compounds that bind the ATX polypeptide can beachieved by isolating the ATX polypeptide/binding complex, andseparating the ATX polypeptide from the binding compound. An additionalstep of characterizing the physical, biological, or biochemicalproperties of the binding compound can also be performed. In oneembodiment, the ATX polypeptide/binding complex can be isolated using aantibody immunospecific for either the ATX polypeptide or the candidatebinding compound. In another embodiment, the complex can be isolatedusing a second binding compound that interacts with either the ATXpolypeptide or the candidate binding compound. In still anotherembodiment, either the polypeptide ATX or the candidate binding compoundcomprises a label or tag that facilitates its isolation, and methods ofthe invention to identify binding compounds include a step of isolatingthe ATX polypeptide/binding complex through interaction with the labelor tag. An exemplary tag of this type is a poly-histidine sequence,generally around six histidine residues, that permits isolation of acompound so labeled using nickel chelation. Other labels and tags, suchas the FLAG tag, thioredoxin, and GST, each of which is well known inthe art.

[0065] An in vitro assay can be performed where the ATX polypeptide canbe immobilized and then contacted with a candidate binding compound. Inan alternative embodiment, the candidate binding compound can beimmobilized and binding of the ATX polypeptide is detected.Immobilization can be accomplished using any of the methods well knownin the art, including covalent bonding to a support, a bead, or achromatographic resin, as well as non-covalent, high affinityinteraction such as antibody binding, or use of streptavidin/biotinbinding wherein the immobilized compound includes a biotin orstreptavidin moiety. Detection of binding can be accomplished, forexample, (i) using a radioactive label on the compound that is notimmobilized, (ii) using of a fluorescent label on the non-immobilizedcompound, (iii) using an antibody immunospecific for the non-immobilizedcompound, (iv) using a label on the non-immobilized compound thatexcites a fluorescent support to which the immobilized compound isattached, as well as other techniques well known in the art.

[0066] A cell based assay that can be used in the method of theinvention for detecting an ATX binding compound is a yeast or mammaliantwo-hybrid assay (Fields and Song, Nature 340:245-246 (1989); Fields,Methods: A Companion to Methods in Enzymoloqy 5:116-124 (1993); U.S.Pat. No. 5,283,173 issued Feb. 1, 1994 to Fields, et al.). Modificationsand variations on the two-hybrid assay have previously been described(Colas and Brent, TIBTECH 16:355-363 (1998)).

[0067] The invention also provides a method for identifying anATX-modulatory compound by measuring the level of an ATX polypeptide inthe presence of a test compound, where a difference in the level of theATX polypeptide in the presence of the test compound compared to in theabsence of the test compound indicating that the test compound is anATX-modulatory compound. In addition, the invention provides a methodfor identifying an ATX-modulatory compound by measuring the level of anATX polypeptide in the presence of a test compound, where a differencein the level of the ATX polypeptide in the presence of the test compoundcompared to in the absence of the test compound indicating that the testcompound is an ATX-modulatory compound, and where the ATX-modulatorycompound is not caffeine or wortmannin. The ATX-modulatory compound candecrease or increase the level of ATX polypeptide.

[0068] Agents that modulate, for example, increase, decrease, or blockthe level of ATX can be identified by incubating a test compound with anATX polypeptide or nucleic acid and determining the effect of the testcompound on ATX activity or expression. The level of ATX can include theexpression level of ATX or an activity level of ATX. The selectivity, orspecificity, of an ATX-modulatory compound can be evaluated by comparingits effects on ATX or an ATX-encoding nucleic acids to its effect onother polypeptides or compounds. Cell based methods, such as two-hybridassays to identify DNAs encoding binding compounds and split hybridassays to identify inhibitors of ATX polypeptide interaction with aknown binding polypeptide, as well as in vitro methods, including assayswhere an ATX polypeptide, ATX-encoding nucleic acid, or a bindingcompound are immobilized, and solution assays are included in thismethod of the invention.

[0069] As understood by those of skill in the art, assay methods foridentifying compounds that modulate an activity generally requirecomparison to a “control.” One type of a control is a reaction or cellthat is treated substantially the same as the test reaction or cellexposed to the compound, with the distinction that the control reactionor cell is not exposed to the compound.

[0070] As disclosed herein, the compounds wortmannin and caffeine canmodulate (inhibit) the level of ATX (Example 3 and Example 7).Wortmannin is known to inhibit ATM kinase and is an irreversibleinhibitor of PIKKs. Caffeine is a known inhibitor of the G2 cell cyclecheckpoint. As disclosed herein, caffeine reversed the accumulation ofG2/M phase cells induced by ATX anti-sense treatment, indicating thatATX deficiency can trigger the activation of a caffeine-sensitive G2checkpoint (Example 7).

[0071] The invention provides a method for identifying an ATX-modulatorycompound where the level of ATX polypeptide is measured by determiningthe kinase activity of the ATX polypeptide. The kinase activity of ATXcan be measured using methods well known in the art such as kinaseassays and immune complex kinase assays as performed herein in Example3. These assays contain ATX, a substrate, and a suitable bufferincluding [g-32 P]ATP and Mn²⁺. Phosphorylated substrates can also bedetected using phospho-specific antibodies.

[0072] In addition, the invention provides a method for identifying anATX-modulatory compound where the level of ATX polypeptide is measuredby determining the phosphorylation of a p53 polypeptide or fragment. Forexample, a GST fusion protein containing the first 70 amino acids of p53(GST-p53₁₋₇₀) can be used as a substrate to measure the level of ATXpolypeptide by its kinase activity (Example 3). In addition to p53, thephosphorylation of hUpf1, a helicase, can be used to measure the levelof ATX polypeptide (Example 3).

[0073] The invention also provides a method for identifying anATX-modulatory compound where the level of ATX polypeptide is measuredby determining the level of p53 polypeptide accumulation. As shownherein, a decrease in ATX polypeptide, such as results from the use ofan anti-sense oligonucleotide, leads to a reduction in p53 polypeptideaccumulation (Example 6). Thus, the level of p53 can be used as ameasure of ATX polypeptide level.

[0074] The invention further provides a method for identifying anATX-modulatory compound where the level of ATX polypeptide is measuredby determining the level of non-sense mediated messenger RNA (mRNA)decay (NMD). NMD is a surveillance mechanism within cells wherebyimperfect mRNAs that contain premature translation termination codonsare preferentially degraded. As disclosed herein, treatment of cellswith an ATX anti-sense oligonucleotide, which reduced endogenous ATXexpression, demonstrated that ATX expression is required for maximal NMDactivity (Example 9). The level of NMD is correlated to the level of ATXin the cell and so the level of NMD can be used as a measure of ATXpolypeptide level.

[0075] ATX-modulatory compounds can be identified that decrease orincrease the level of ATX polypeptide or nucleic acid. A decrease in thelevel of ATX can be a partial reduction or a total blockage of the levelof ATX, and an increase in the level of ATX can be a partial increase oran induction of the level of ATX from a previously undetectable level.For example, an ATX-modulatory compound can increase the level of NMDactivity in a cell. It can be desirable to increase the level of NMDactivity in a cell in order to protect the cell from deleteriousgain-of-function mutations caused by truncated polypeptides resultingfrom the translation of imperfect. mRNAs that contain prematuretranslation termination. Alternatively, an ATX-modulatory compound candecrease the level of NMD activity in a cell. It can be desirable todecrease the level of NMD activity in a cell in some cases where thetruncated polypeptide does not have a deleterious effect but insteadretains some activity that can compensate for the normal gene function.

[0076] ATX-modulatory compounds can include, for example, antibodies andother proteins or peptides which specifically bind to an ATX polypeptideor an ATX-encoding nucleic acid, oligonucleotides which bind to an ATXpolypeptide or an ATX gene sequence, and other non-peptide compounds,for example, isolated or synthetic organic and inorganic molecules,which specifically react with an ATX polypeptide or underlying nucleicacid. ATX-modulatory compounds of the invention can interactspecifically or exclusively to an ATX polypeptide or ATX-encodingnucleic acid, however, modulators that interact with an ATX polypeptideor an ATX-encoding nucleic acid with higher affinity or avidity comparedto other compounds are also included in the invention. Mutant ATXpolypeptides which affect the enzymatic activity or cellularlocalization of the wild-type ATX polypeptides are also contemplated bythe invention. Targets for the development of ATX-modulatory compoundscan include, for example: (1) regions of an ATX polypeptide whichcontact other proteins, (2) regions that localize an ATX polypeptidewithin a cell, (3) regions of an ATX polypeptide which bind substrate,(4) allosteric regulatory binding site(s) of an ATX polypeptide, (5)phosphorylation site(s) of an ATX polypeptide as well as other regionsof the protein where covalent modification regulates biological activityand (6) regions of an ATX polypeptide which are involved inmultimerization of subunits. Still other ATX-modulatory compoundsinclude those that recognize specific ATX-encoding and regulatorynucleic acid sequences. ATX-modulatory compounds that modulate the levelof ATX can be therapeutically useful in treatment of diseases andphysiological conditions in which ATX is known or suspected to beinvolved.

[0077] Methods of the invention to identify ATX-modulatory compoundsinclude variations on any of the methods described above to identify ATXbinding compounds, the variations including techniques where a bindingcompound has been identified and the binding assay is carried out in thepresence and absence of a candidate ATX-modulatory compound. Amodulatory compound is identified in those instances where the level ofbinding between an ATX polypeptide and a binding compound changes in thepresence of the candidate modulatory compound compared to the level ofbinding in the absence of the candidate modulatory compound. AnATX-modulatory compound that increases binding between an ATXpolypeptide and the binding compound is described as an enhancer oractivator, and a modulatory compound that decreases binding between theATX polypeptide and the binding compound is described as an inhibitor.In vitro methods of the invention are amenable to high throughput assaysas described below.

[0078] In addition to the assays described above which can be modifiedto identify binding compounds, other methods are contemplated toidentify modulatory compounds. In one embodiment, methods of theinvention can include use of the split hybrid assay as generallydescribed in WO98/13502 and variations on this method as described inWO95/20652.

[0079] The methods of the invention can also utilize high throughputscreening (HTS) assays to identify compounds that interact with orinhibit biological activity of an ATX polypeptide. HTS assays permitscreening of large numbers of compounds in an efficient manner.Cell-based HTS systems include melanophore assays, yeast-based assaysystems, and mammalian cell expression systems (Jayawickreme and Kost,Curr. Opin. Biotechnol. 8:629-634 (1997)). Automated (robotic) andminiaturized HTS assays are also embraced (Houston and Banks, Curr.Opin. Biotechnol. 8:734-740 (1997)). HTS assays are designed to identify“hits” or “lead compounds” having the desired property, from whichmodifications can be designed to improve the desired property. Chemicalmodification of the “hit” or “lead compound” is often based on anidentifiable structure/activity relationship (SAR) between the “hit” andthe ATX polypeptide.

[0080] There are a number of different libraries used for theidentification of small molecule modulators, including, (1) chemicallibraries, (2) natural product libraries, and (3) combinatoriallibraries comprised of random peptides, oligonucleotides or organicmolecules.

[0081] Chemical libraries consist of structural analogs of knowncompounds or compounds that are identified as “hits” or “leads” vianatural product screening. Natural product libraries are collectionsfrom microorganisms, animals, plants, or marine organisms which are usedto create mixtures for screening by, for example, (1) fermentation andextraction of broths from soil, plant or marine microorganisms or (2)extraction of plants or marine organisms. Natural product librariesinclude polyketides, non-ribosomal peptides, and variants (non-naturallyoccurring) variants thereof. Combinatorial libraries are composed oflarge numbers of peptides, oligonucleotides or organic compounds as amixture. They can be prepared by traditional automated synthesismethods, PCR, cloning or proprietary synthetic methods. Libraries thatcan be utilized by the invention include peptide and oligonucleotidecombinatorial libraries. Still other libraries of interest includeprotein, peptidomimetic, multiparallel synthetic collection,recombinatorial, and polypeptide libraries. For a review ofcombinatorial chemistry and libraries created therefrom, see Myers,Curr. Opin. Biotechnol. 8:701-707 (1997). Identification of modulatorsthrough use of the various libraries described herein permitsmodification of the candidate “hit” (or “lead”) to optimize the capacityof the “hit” to modulate activity.

[0082] Anti-sense oligonucleotides which recognize and hybridize tonucleic acid encoding ATX can also be utilized by the methods of theinvention. Full length and fragment anti-sense oligonucleotides areprovided. One skilled in the art of will appreciate that fragmentanti-sense molecules of the invention include (i) those whichspecifically or exclusively recognize and hybridize to ATX-encoding RNA(as determined by sequence comparison of DNA encoding ATX to DNAencoding other molecules) as well as (ii) those which recognize andhybridize to RNA encoding variants of the ATX family of proteins.Antisense oligonucleotides that hybridize to RNA encoding other membersof the PIKK family of proteins are also identifiable through sequencecomparison to identify characteristic, or signature, sequences for thefamily of molecules. Identification of sequences unique to ATX-encodingnucleic acids, as well as sequences common to the family ofPIKK-encoding nucleic acids, can be deduced through use of any publiclyavailable sequence database, or through use of commercially availablesequence comparison programs. After identification of the desiredsequences, isolation through restriction digestion or amplificationusing any of the various polymerase chain reaction techniques well knownin the art can be performed. Anti-sense oligonucleotides can be used forregulating expression of ATX by those cells expressing ATX mRNA.Antisense molecules are generally from about 5 to about 100 nucleotidein length, and preferably are about 10 to 20 nucleotides in length.Antisense nucleic acids capable of specifically binding to ATXexpression control sequences or ATX RNA are introduced into cells, forexample, by a viral vector or colloidal dispersion system such as aliposome.

[0083] The anti-sense nucleic acid binds to the ATX-encoding targetnucleotide sequence in the cell and prevents transcription ortranslation of the target sequence. Phosphorothioate andmethylphosphonate anti-sense oligonucleotides are specificallycontemplated for therapeutic use by the invention. The anti-senseoligonucleotides may be further modified by poly-L-lysine, transferrinpolylysine, or cholesterol moieties at their 5′ end.

[0084] The invention also provides methods to modulate ATX expressionthrough the use of RNA interference (RNAi) (Brummelkamp et al., Science296:550-553 (2002); Elbashir et al., Nature 411:494-498 (2002)). RNAi isa process of sequence-specific gene silencing by post-transcriptionalRNA degradation, which is initiated by double-stranded RNA (dsRNA)homologous in sequence to the silenced gene. A double-stranded RNA(dsRNA) that is used for RNAi is referred to herein as an “interferingRNA.” For example, a suitable dsRNA for RNAi can contain sense andantisense strands of about 21 contiguous nucleotides corresponding tothe gene to be targeted that form 19 RNA base pairs, leaving overhangsof two nucleotides at each 3′ end (Elbashir et al., supra; Bass, Nature411:428-429 (2001); Zamore, Nat. Struct. Biol. 8:746-750 (2001)). dsRNAsof about 25-30 nucleotides have also been used successfully for RNAi(Karabinos et al., Proc. Natl. Acad. Sci. 98:7863-7868 (2001). dsRNA canbe synthesized in vitro and introduced into a cell by methods known inthe art. By using RNAi methods, the targeted RNA is degraded, andtranslation of the target polypeptide is decreased or abolished.

[0085] The invention further provides methods to modulate ATX expressionthrough the use of ribozymes (Gibson and Shillitoe, Mol. Biotech.7:125-137 (1997)). Ribozyme technology can be utilized to inhibittranslation of ATX mRNA in a sequence specific manner through (i) thehybridization of a complementary RNA to a target mRNA and (ii) cleavageof the hybridized mRNA through nuclease activity inherent to thecomplementary strand. Ribozymes can be identified by empirical methodsor be specifically designed based on accessible sites on the target mRNA(Bramlage, et al., Trends in Biotech 16:434-438 (1998)). Delivery ofribozymes to target cells can be accomplished using either exogenous orendogenous delivery techniques well known in the art. Exogenous deliverymethods can include use of targeting liposomes or direct localinjection. Endogenous methods include use of viral vectors and non-viralplasmids. Ribozymes can be ATX-modulatory compounds and specificallymodulate expression of ATX when designed to be complementary to regionsunique to a nucleic acid encoding ATX. Specifically modulate means thatribozymes of the invention exclusively recognize a nucleic acid encodingATX.

[0086] The invention further provides methods to modulate transcriptionof ATX through use of oligonucleotide-directed triple helix formation(Lavrovsky, et al., Biochem. Mol. Med. 62:11-22 (1997)). Triple helixformation is accomplished using sequence specific oligonucleotides whichhybridize to double stranded DNA in the major groove as defined in theWatson-Crick model. Hybridization of a sequence specific oligonucleotidecan thereafter modulate activity of DNA-binding proteins, including, forexample, transcription factors and polymerases. Target sequences forhybridization include promoter and enhancer regions to permittranscriptional regulation of ATX expression. In addition to use ofoligonucleotides, triple helix formation techniques of the inventionalso include use of peptide nucleic acids as described in Corey, TIBTECH15:224-229 (1997). Oligonucleotides which are capable of triple helixformation are also useful for site-specific covalent modification oftarget DNA sequences. Oligonucleotides useful for covalent modificationcan be coupled to various DNA damaging agents as described in Lavrovsky,et al. (supra).

[0087] Mutations in the ATX gene can result in loss of normal functionof the ATX gene product and underlie ATX-related human disease states.The invention therefore provides gene therapy methods to restore ATXactivity in treating those disease states described herein. Delivery ofa functional ATX gene to appropriate cells is effected ex vivo, in situ,or in vivo by use of vectors, for example, viral vectors such asadenovirus, adeno-associated virus, or a retrovirus, or ex vivo by useof physical DNA transfer methods such as liposomes or chemicaltreatments (Anderson, Nature, supplement to vol. 392, no. 6679, pp.25-20(1998)). Alternatively, in some human disease states, preventing theexpression of, or inhibiting the activity of, ATX can be useful intreating the disease states. In this case, anti-sense therapy or genetherapy, for example, where a dominant negative ATX mutant is introducedinto a target cell type, can be applied to negatively regulate theexpression of ATX.

[0088] The invention provides a method for modulating cell survival byintroducing an ATX-modulatory compound identified by the methodsdescribed above into a cell in an amount effective to modulate survivalof the cell. For example, the ATX-modulatory compound can decrease orincrease cell survival.

[0089] A level of cell death or cell survival can be measured by any ofa variety of methods known to one skilled in the art. For example,trypan blue staining can be used to measure the level of cell death in acell. In addition, clonogenic assays, as described herein, can be used(Example 5). Other staining methods, for example, propidium iodide andAlomar Blue, also can be used to measure cell death. The stained cellscan be visualized in any way that is convenient, for example, bymicroscopy or flow cytometry (FACS). In addition, cell viability andcell proliferation assays such as the lactose dehydrogenase (LDH) assayand the MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliumbromide) assay are commercially available and can be used to measurecell viability. In addition, the uptake of 3H thymidine can be used toaccess the viability of cells.

[0090] The invention further provides a method for modulating cellsurvival by introducing an ATX-modulatory compound into a cell where thecell is exposed to a stressor agent. As described further above, astressor agent is any agent that can induce a stress response pathwaywithin a cell. A stressor agent can include, for example, UV light,ionizing radiation, reactive oxygen intermediates, or a chemical agentsuch as a cytotoxic or chemotherapeutic agent. In addition,environmental conditions such as excessive heat can induce a stressresponse pathway within a cell resulting in, for example, the inductionof heat shock proteins. Stress response pathways include DNA repairpathways, non-sense mediated mRNA decay (NMD), heat shock pathways, theinduction of apoptosis, activation of the NFkB transcription factor,activation of the stress-activated MAP kinase pathways including, forexample, JNK and p38 pathways, and activation of ubiquitin-dependentproteolysis.

[0091] An example of an ATX-modulatory compound of the invention is anantisense oligonucleotide. The invention provides a method fordecreasing cell survival by introducing an antisense oligonucleotide,such as SEQ ID NO: 11 into a cell in an amount effective to decreasesurvival of the cell.

[0092] Association of ATX with cell cycle progression makes compositionsof the invention, including for example an ATX polypeptide, an inhibitorthereof, an antibody, or other modulator of ATX expression or biologicalactivity, useful for treating a number of conditions. For example, theinvention provides a method for treating a condition characterized byexcessive cell survival or cell growth by administering to a patienthaving such a condition an effective amount of an ATX-modulatorycompound where the effective amount of the compound increases celldeath. For example, an ATX-modultory compound can be given to a patientwith a neoplastic condition.

[0093] An ATX-modulatory compound that decreases the level of ATX canenhance the radiosensitivity or chemosensitivity of neoplastic cells.Therefore, it is contemplated that an ATX-modulatory compound can begiven alone or in combination with another agent such as a cytotoxic orchemotherapeutic agent. Several cytotoxic agents, such as radiation, andchemotherapeutic agents, such as cis-platin, are well known in the art.An appropriate agent can be chosen based on several factors, such as theparticular type of neoplastic condition at issue or the ability of thepatient to tolerate the agent. For example, focused radiation therapy,including brachytherapy, can be used in conjunction with an ATXinhibitory compound in order to induce tumor cell death while minimizingcytotoxic effects on normal tissue.

[0094] A “neoplastic condition,” refers to a condition associated withhyperproliferation of cells and includes benign and malignant expandinglesions of proliferating cells. Neoplastic conditions include benign andmalignant hyperproliferative disorders. A benign neoplasm grows in anexpansile manner, displacing or compressing surrounding tissues ratherthan invading them. A malignant neoplasm refers to a large group ofdiseases characterized by uncontrolled growth and spread of abnormalcells. Cancer, for example, is a malignant neoplastic condition thatencompasses many sub-conditions that are characterized by insufficientdeath of abnormal cells. Tumors of the colon, prostate, lung, cervix,stomach, breast and skin are examples of neoplastic conditions.

[0095] Aberrant ATX activity can be associated with various forms ofcancer in, for example, adult and pediatric oncology, including growthof solid tumors/malignancies, myxiod and round cell carcinoma, locallyadvanced tumors, metastatic cancer, human soft tissue sarcomas, cancermetastases, including lymphatic metastases, squamous cell carcinoma ofthe head and neck, esophageal squamous cell carcinoma, oral carcinoma,blood cell malignancies, including multiple myeloma, leukemias, effusionlymphomas (body cavity based lymphomas), thymic lymphoma lung cancer,including small cell carcinoma, non-small cell cancers, breast cancer,including small cell carcinoma and ductal carcinoma, gastrointestinalcancers, including stomach cancer, colon cancer, colorectal cancer,polyps associated with colorectal neoplasia, pancreatic cancer, livercancer, urological cancers, including bladder cancer, including primarysuperficial bladder tumors, invasive transitional cell carcinoma of thebladder, and muscle-invasive bladder cancer, prostate cancer,malignancies of the female genital tract, including ovarian carcinoma,primary peritoneal epithelial neoplasms, cervical carcinoma, uterineendometrial cancers, and solid tumors in the ovarian follicle, kidneycancer, including renal cell carcinoma, brain cancer, includingintrinsic brain tumors, neuroblastoma, astrocytic brain tumors, gliomas,metastatic tumor cell invasion in the central nervous system, bonecancers, including osteomas, skin cancers, including malignant melanoma,tumor progression of human skin keratinocytes, and squamous cell cancer,hemangiopericytoma, and Kaposi's sarcoma.

[0096] Aberrant ATX activity also can be associated with otherconditions which include aberrant apoptotic mechanisms, includingabnormal caspase activity; aberrant enzyme activity associated with cellcycle progression, including for example cyclins A, B, D and E;alterations in viral (such as Epstein-Barr virus, papillomavirus)replication in latently infected cells; chromosome structureabnormalities, including genomic stability in general, unrepairedchromosome damage, telomere erosion (and telomerase activity), breakagesyndromes including for example, Sjogren's syndrome and Nijimegenbreakage syndrome; embryonic stem cell lethality; abnormal embyonicdevelopment; sensitivity to ionizing radiation; acute immune complexalveolitis; and Fanconi anemia. ATX-modulatory compounds can be usedalone or in combination with another agent in the treatment of theseconditions.

[0097] The invention also provides a method for treating a conditioncharacterized by excessive cell death by administering to a patienthaving such a condition an effective amount of an ATX-modulatorycompound where the effective amount of the compound increases cellsurvival. For example, an ATX-modultory compound can be given to apatient with a neurodegnerative condition in order to increase neuronalcell survival. In addition the invention provides a method of prolongingthe in vivo survival of transplanted cells for the treatment of adisease or pathological condition. Also, for example, a compound thatincreases the level of ATX can be given to a patient who is exposed tostressors such as UV light in order to protect against geneticmutations.

[0098] The effective compounds of the invention described herein canoptionally be formulated together with a pharmaceutically acceptablecarrier for delivery to a cultured cell or to a subject. Suitablepharmaceutically acceptable carriers are well known in the art andinclude, for example, aqueous or organic solvents such asphysiologically buffered saline, glycols, glycerol, oils or injectableorganic esters. A pharmaceutically acceptable carrier can also contain aphysiologically acceptable compound that acts, for example, to stabilizeor increase the solubility of a pharmaceutical composition. Such aphysiologically acceptable compound can be, for example, a carbohydrate,such as glucose, sucrose or dextrans; an antioxidant, such as ascorbicacid or glutathione; a chelating agent; a low molecular weightpolypeptide; or another stabilizer or excipient. Pharmaceuticallyacceptable carriers, including solvents, stabilizers, solubilizers andpreservatives, are described, for example, in Martin, Remington's Pharm.Sci., 15th Ed. (Mack Publ. Co., Easton, 1975).

[0099] Those skilled in the art can formulate the therapeutic moleculesto ensure proper distribution in vivo. For example, the blood-brainbarrier (BBB) excludes many highly hydrophilic compounds. To ensure thatthe effective compounds of the invention cross the BBB, if desired, theycan be formulated, for example, in liposomes, or chemically derivatized.For a review of strategies for increasing bioavailability of polypeptidedrugs in the brain, and of methods for determining the permeability ofpolypeptides through the BBB using in vitro and in vivo assays, seeEngleton et al., Peptides 9:1431-1439 (1997). Strategies that have beensuccessfully used to increase the permeability of other neuropeptidesthrough the BBB are particularly contemplated. Modifications to apolypeptide of the invention that can increase its BBB penetrationinclude conjugating the peptide to a lipophilic moiety, such as alipophilic amino acid or methyldihydropyridine. Similar modifications toinvention polypeptides or peptidomimetics are likewise expected to beadvantageous.

[0100] Methods of ensuring appropriate distribution in vivo can also beprovided by rechargeable or biodegradable devices, particularly wheregradients of concentrations of drug in a tissue are desired. Variousslow release polymeric devices are known in the art for the controlleddelivery of drugs, and include both biodegradable and non-degradablepolymers and hydrogels. Those skilled in the art understand that thechoice of the pharmaceutical formulation and the appropriate preparationof the composition will depend on the intended use and mode ofadministration.

[0101] The effective compounds of the invention can be administered to asubject by any effective route. Suitable routes for delivering thetherapeutic molecules of the invention include topically, intraocularly,intradermally, parenterally, orally, intranasally, intravenously,intramuscularly, intraspinally, intracerebrally and subcutaneously. Thepresent invention also provides compounds containing an acceptablecarrier such as any of the standard pharmaceutical carriers, includingphosphate buffered saline solution, water and emulsions such as an oiland water emulsion, and various types of wetting agents.

[0102] An effective dose of an effective compound of the invention canbe determined, for example, by extrapolation from the concentrationrequired in the ATX binding or ATX activity assays described herein; orfrom the dose required to modulate cell proliferation. An effective doseof an effective compound of the invention for the treatment of apathology can also be determined from appropriate animal models, such astransgenic mice. Animal models for pathologies such as tumors arewell-known in the art. An effective dose for treating this disease is adose that results in either partial or complete regression of the tumor,reduction in metastasis, reduced discomfort, or prolonged life span. Theappropriate dose for treatment of a human subject with a therapeuticmolecule of the invention can be determined by those skilled in the art,and is dependent on the nature and bioactivity of the particularcompound, the desired route of administration, the gender, age andhealth of the individual, the number of doses and duration of treatment,and the particular condition being treated.

[0103] It is understood that modifications which do not substantiallyaffect the activity of the various embodiments of this invention arealso included within the definition of the invention provided herein.Accordingly, the following examples are intended to illustrate but notlimit the present invention.

EXAMPLE 1 Molecular Cloning of ATX

[0104] This example shows the cloning of ATX nucleic acids. During aBLAST search for mTOR-related proteins, it was noted that an expressedsequence tag (EST) (KIAA0421) contained a 5′-terminus with an openreading frame (ORF) that bore clear homology to a conserved region inthe catalytic domains of PIKK family members. To access the full-lengthcDNA, the EST was used to generate a primer for 5′-RACE with human braincDNA as the template. The initial 5′-RACE product extended the region ofhomology with the PIKK catalytic domain. Sequential screens of humanbrain (Clontech #HL1128a) and Jurkat T cell cDNA libraries (Stratagene#936219), combined with 5′-RACE of brain and Jurkat cDNA, resulted inthe isolation of several overlapping DNA fragments that were assembledinto approximately 12 kb of contiguous nucleotide sequence. This cDNAcontains an ORF of 10,563 nucleotides with an additional 1.8 kb of3′-UTR, and encodes a 3,521 amino acid polypeptide with a deducedmolecular mass of 395 kDa. The first nucleotide of the ATG translationinitiation codon in exon 6 as has been designated as nucleotide “1”, andnucleotides upstream of this ATG are identified in the 3′ to 5′direction with negative numbers. The conclusion that this. sequence wasderived from a single mRNA transcript was confirmed by PCR with primersthat were complementary to the extreme 5′-terminus (nucleotides −90 to−67) and 3′-terminus (nucleotides 10,553 to 10,570) of the correspondingcDNA. The cloned cDNA sequence is contained in a genomic BAC clone(AC020716), which allowed localization of the gene encoding thisputative PIKK family member to human chromosome 16. Based on itsfunctional overlap with ATM, this new PIKK family member was named“ATX”.

[0105] The collective results of the 5′-RACE and RT-PCR assays of mRNAderived from Jurkat T cell, human brain, and other human cell linesindicated that the ATX locus gives rise to several mRNA transcripts(FIG. 1A). One repetitively isolated ATX cDNA clone contains exon 4spliced directly to exon 6, and yields the 3,521 amino acid polypeptidedescribed above. This mRNA transcript and encoded polypeptide has beendesignated “long ATX”, to distinguish it from a “short ATX” polypeptide(3,031 amino acids) produced as a result of allelic variation in exon15, which leads to the insertion of 27 nucleotides beginning atnucleotide 1427 (FIG. 1A). This sequence alteration causes the insertionof two in-frame stop codons, and use of the next available ATG codon (inexon 16) as the translational start site gives rise to theamino-terminally truncated, short form of ATX. The 5′ end of the ATXallele that encodes short ATX is contained within a second genomic BACclone (AC003007) derived from human chromosome 16. Yamashita et al. haveidentified two human cDNA clones, both designated hSMG-1, one of which(FIG. 1A, second from bottom, SEQ ID NO: 7) was similar to the long ATXcDNA clone (SEQ ID NO:1) (Yamashita et al., Genes and Development15:2215-2228 (2001)). Exon 5 was-not included in our long cDNA clone dueto the infrequent appearance of this exon during our 5′-RACE and RT-PCRanalyses of human cell line-derived mRNA. Furthermore, the minority ofcDNAs that did include the exon 5 sequence frequently contained exon 3,which placed an in-frame stop codon at the 5′-end of this cDNA (FIG.1A). The longest ATX cDNA clone (ORF beginning at exon 2, SEQ ID NO:9)identified by Yamashita et al. (Yamashita et al., supra, 2001) was alsoisolated in our screening procedure. However, it was repeatedly foundthat the exon 3-associated stop codon was present in this transcript.

EXAMPLE 2 Expression of Endogenous and Recombinant ATX

[0106] In order to examine the expression of ATX mRNA in varioustissues, a multiple tissue Northern blot was hybridized with a32P-labelled, ATX cDNA probe that spanned exons 38-39(nucleotides5,071-5,370). The ATX probe detected a major mRNA species that, based onits electrophoretic mobility, was significantly larger the 9.5 kbcalibration marker, and could reasonably accommodate the predicted ORF(10.5 kb) of long ATX (data not shown). This ATX transcript was widelyexpressed in human tissues, with the highest levels observed in heartand skeletal muscle. These results are consistent with those obtained inimmunoblot analyses with ATX-specific antibodies, which showed that ATXprotein was uniformly expressed in hematopoietic, mesenchymal, andepithelial cell lines (data not shown). Database searches with the ATXamino acid sequence revealed the highest degree of homology to C.elegans SMG1, a protein required for NMD in the worm. Both ATX andCeSMG1 contain the PI 3-kinase related catalytic domain, whichidentifies these proteins as members of the PIKK family (FIG. 1B).Outside of the catalytic domain, the regional sequence homology betweenATX and other PIKK family members was limited to the FKBP-12rapamycinbinding (FRB, designated NH2 in FIG. 1B) domain of mTOR, and to the NH1and NH2 domains of CeSMG1. The FRB domain mediates the high-affinityinteraction between mTOR and the antiproliferative FKBP12rapamycincomplex (Chen et al., Proc. Natl. Acad. Sci. USA, 92:4947-4951 (1995)).However, the FRB-related domain of ATX does not confer any detectablebinding affinity for FKBP12rapamycin (data not shown); hence, it isunlikely that ATX is a relevant target for rapamycin in intact cells.The expression of the short and long forms of ATX were compared aftertransient transfection of the respective cDNAs into human embryonickidney 293T cells. The short ATX polypeptide was poorly expressedrelative to long ATX (data not shown). However, these results do notexclude the possibility that the shorter form of ATX is expressed andcontributes to the overall functions of ATX in mammalian cells.

[0107] In order to compare the translation product derived from the longATX cDNA with the endogenously expressed ATX polypeptide, HEK 293T cellswere transfected with HA-tagged expression plasmids encoding eitherwild-type ATX (HA-ATXWT) or a catalytically inactive ATX mutant(HA-ATXKI). The HA-ATXKI mutant contains an Asp>Ala substitution at aconserved residue (Asp-2195) in the ATX catalytic domain. Similarmutations in the catalytic domains of ATM, ATR, and DNA-PKCS have beenshown to generate kinase-inactive (KI) versions of these PIKK familymembers (Canman et al., Science, 281:1677-1679 (1998); Cliby et al.,EMBO Journal, 17:159-169 (1998); Hunter, supra, 1995).

[0108] For analyses of the endogenous ATX protein, two different rabbitpolyclonal antibodies were prepared against GST fusion proteinscontaining peptide fragments derived from ATX. The first antibody (α-ATXAb-1) was generated against a GST fusion protein containing amino acids2281-2339 of ATX, while the second (α-ATX Ab-2) was obtained fromimmunizations with GST fused to amino acids 1691-1790 of ATX. The α-ATXAb1 immunoblot analyses of whole cell extracts from non-transfected orHA-ATX-transfected HEK 293T cells revealed a single major immunoreactiveband migrating at the predicted molecular mass of ˜395 kDa (data notshown). An immunoreactive protein bearing a nearly identicalelectrophoretic mobility was detected in α-HA immunoprecipitates fromtransfected 293T cells. These results indicate that the molecular massof the recombinant protein produced from the long ATX cDNA correspondsclosely to that of the endogenous ATX protein. Consistent with thepredicted size of ATX, the band recognized by the α-ATX antibodiesmigrated with a significantly lower electrophoretic mobility than eitherATM (molecular mass, 370 kDa) or ATR (molecular mass, 305 kDa).

[0109] Methods:

[0110] Cloning

[0111] The longer ATX ORF was appended with an amino-terminalhemagglutinin (HA) epitope tag sequence (CYPYDVPDYASL), and wassubsequently amplified as two partially overlapping fragments fromJurkat cDNA. The nucleotide at position 4,620 was mutated in each of thetwo PCR products to create a SacII site that could be utilized to ligatethe two ATX fragments, which were inserted into the XhoI and NotI sitesof pcDNA3.1 (Invitrogen) (HA-ATX). The mutation used to generate theSacII did not alter the ATX polypeptide sequence. The expression vectorencoding the catalytically inactive ATX mutant (HA-ATXKI) contains anAla substitution at Asp-2195, which was generated by site-directedmutagenesis with the QuickChange kit (Stratagene). All plasmidconstructs were sequenced to insure the fidelity of the PCR and cloningprocedures.

[0112] Cell lines

[0113] U2OS osterosarcoma and human embryonic kidney (HEK) 293T cellswere cultured in low-glucose Dulbecco's Modified Eagle's Medium (DMEM),supplemented with 10% fetal bovine serum. The ATM-deficient humanfibroblast line, AT4BI, was maintained in DMEM/F-12 medium supplementedwith 10% fetal bovine serum. Where indicated, cells were γ-irradiatedwith a 137Cs source or UV irradiated with a UV-B source (λmax, 305 nm).

[0114] Antibodies

[0115] ATX-specific antibodies were raised by immunizing rabbits(Cocalico Biologicals, Inc.) with the indicated glutathioneS-transferase (GST) fusion protein. Anti-ATX Ab-1 was raised against aGST fusion protein containing ATX amino acids 2281-2339, and α-ATX Ab-2was raised against a GST fusion protein containing ATX amino acids1691-1790. For purification of α-ATX Ab-2, GST-reactive antibodies werefirst absorbed on GSH-agarose. The flow-through fraction was thenaffinity purified over Affi-Gel 15 (BIO-RAD) coupled to theGST-ATX1691-1790 fusion protein. The α-PLC-γ1 antiserum was prepared asdescribed (Secrist et al., J. Biol. Chem., 268: 5886-5893 (1993)). Theα-ATM (Ab-3), α-ATR (Ab-1), α-phospho-Ser15-p53, and α-p53 (Ab-6)reagents were obtained from Oncogene Science Research Products.Additional antibodies were obtained from (sources in parentheses): α-HA(clone 12CA5; BabCo), α-FLAG-M2 and α-tubulin (Sigma), α-Cds1/Chk2(Upstate Biotechnology), and α-GAL4 (clone RK5C1; Santa CruzBiotechnology).

[0116] Two-dimensional PAGE

[0117] HEK 293 cells were lysed and protein analyzed as described {Pal,2001 #1360}, except that cellular extracts were incubated for 2 h withα-FLAG-M2 mAb, followed by 2 h with protein G agarose (Sigma) toimmunoprecipitate the FLAG-hUpf1 protein. Prior to elution, theimmunoprecipitates were washed in lysis buffer as described {Pal et al.,Rna 7:5-15 (2001) #1360}.

[0118] Immunofluorescence

[0119] For immunofluorescence microscopy of endogenous ATX, 6×104 U2OScells were plated onto 22-mm2 glass coverslips. After 40 h, cells wereexposed to 400 J/m2 UV-B, then fixed 1, 4 or 8 hrs later inphosphate-buffered saline (PBS) containing 4% paraformaldehyde for 20min, and incubated in methanol at −20° for 15 min. The coverslips wererehydrated in phosphate-buffered saline (PBS) and incubated overnight at40° C. in blocking solution (PBS containing 3% BSA and 2% goat serum).Coverslips were subsequently overlayed for 1 h with affinity purifiedα-ATX Ab-2 (1 μg per ml) in blocking solution at room temperature.Coverslips were washed with PBS, 0.2% Tween-20, and overlayed for 45 minat room temperature with fluorescein isothiocyanate (FITC)-conjugatedgoat anti-rabbit IgG (Caltag) (1:2000 in blocking solution). Sampleswere then washed and incubated with 100 μg per ml RNaseA in PBS for 30min, followed by 1 μg per ml propidium iodide for 5 min. After extensivewashing in PBS containing 0.2% Tween-20, coverslips were mounted onslides with an aqueous anti-fade mounting reagent (Vectashield, VectorLaboratories). Immunofluorescence images were generated with a CarlZeiss LSM410 scanning laser confocal microscope.

EXAMPLE 3 Characterization of ATX Kinase Activity

[0120] With the exception of the TOR proteins, the PIKK family membersthat bear functional catalytic domains phosphorylate substrates bearingthe Ser/Thr-Gln motif (Tibbetts and Abraham, supra, 2000). To determinewhether the ATX kinase domain displayed a similar phosphorylation siteselectivity, HEK 293T cells were transfected with a plasmid vectorencoding HA-tagged ATXWT, ATXKI, or, for comparative purposes, HA-ATMWT.Detergent extracts from the transfected cell populations wereimmunoprecipitated with α-HA antibody, and protein kinase assays wereperformed in buffer containing Mn2+, [γ-32P]ATP, and a GST fusionprotein containing the first 70 amino acids of p53 (GST-p53₁₋₇₀) as thesubstrate (FIG. 1C) The GST-p53₁₋₇₀ protein was previously identified asa substrate for ATM and ATR in immune complex kinase assays (Tibbetts etal., Genes and Development 13:152-157 (1999)). Interestingly, thespecific kinase activity of HA-ATXWT towards GST-p53₁₋₇₀ wassignificantly higher than that of HA-ATM (FIG. 1C, left panel). Based onthe ratios of 32P incorporation into substrate to levels of HA-taggedprotein kinase, it can be estimated that the specific kinase activity ofATXWT was approximately 3.5-fold higher than that of ATMWT. As observedwith ATM as the test kinase (Banin et al., Science 281:1674-1677 (1998);Canman et al., Science 281:1677-1679 (1998); Tibbetts et al., supra,1999), phosphorylation of GST-p53₁₋₇₀ by ATXWT was nearly abolished bysubstitution of the Ser-15 residue in p53 with Ala (FIG. 1C, rightpanel). Because Ser-15 resides in the optimal sequence (LSQE) forphosphorylation by ATM (O'Neill et al., J. Biol. Chem. 275:22719-22727(2000)), this finding indicates that ATX is a Ser/Thr-Gln-directedkinase, with the potential to phosphorylate a set of substrates thatoverlaps with those modified by ATM. In contrast to the amino-terminalfragment of p53, the PHAS-I/4E-BP1 protein, which is the prototypicalsubstrate for mTOR, was poorly phosphorylated by HA-ATXWT in immunecomplex kinase assays (data not shown).

[0121] The amino acid sequences surrounding the four phosphorylationsites (LSQP, LSQD, LSQD, and LSQY) identified in this study resemble theconsensus site for phosphorylation by ATM (O'Neill et al., supra, 2000).A GST fusion protein that contained the carboxyl-terminal region ofhUpf1 (amino acids 1019-1118), including all four of the ATXphosphorylation sites was constructed. This GST-hUpf11019-1118 proteinwas tested as a substrate for HA-ATXWT versus HA-ATMWT in immune complexkinase assays. Once again, this substrate was phosphorylated by both ATMand ATX, with the latter protein kinase showing the higher specificcatalytic activity under these in vitro assay conditions (FIG. 1C, leftpanel). Furthermore, the results of repeated assays indicated thatGST-p53₁₋₇₀ was more avidly phosphorylated by ATX than was theGST-hUpf11019-1118 substrate.

[0122] The protein kinase activities of the mammalian PIKKscharacteristically display a strong dependence on Mn2+ as a cofactor forthe phosphotransferase reaction, and variable sensitivity to inhibitionby wortmannin (Abraham, Genes and Development 15:2177-2196 (2001)). Inour studies, the protein kinase activity of ATX was also dependent onthe addition of Mn2+ to the kinase reaction buffer (data not shown). Inaddition, pretreatment of the immunoprecipitated HA-ATXWT protein withwortmannin resulted in a concentration-dependent inhibition ofGST-p53₁₋₇₀ phosphorylation. The drug concentration required for 50%inhibition (IC50) of ATX activity in vitro was between 10 and 100 nM(FIG. 1D), which is comparable to the previously published IC50 (80 nM)for wortmannin as an ATM inhibitor (Sarkaria et al., Cancer Res.58:4375-4382 (1998)). Wortmannin is an irreversible inhibitor of PIKKs(Walker et al., Molecular Cells 6:909-919 (2000)) and can be used toassess the potency of this drug as an ATX inhibitor in intact cells. Tothis end, U2OS osteosarcoma cells were pretreated for 30 min with theindicated concentrations of wortmannin, followed by the preparation ofcellular extracts for immunoprecipitation of endogenous ATX with α-ATXAb-2. Under these conditions, wortmannin inhibited ATX kinase activitywith an IC50 of 1-3 μM; which is considerably higher than that observedfollowing direct treatment of the immunoprecipitated protein kinase withthis drug (FIG. 1D). A similarly dramatic decrease in the inhibitorypotency of wortmannin was observed with ATM as the target enzyme inintact cells (Sarkaria et al., supra, 1998).

[0123] Immune Complex Kinase Assays

[0124] Native or recombinant ATX proteins were immunoprecipitated fromcell extracts as described above, and the immunoprecipitates were washedtwice in lysis buffer, once in high-salt buffer (100 mM Tris-HCl, pH7.4, 500 mM LiCl) and once in kinase buffer (10 mM Hepes, pH 7.4, 50 mMNaCl, 50 mM β-glycerol phosphate). Forty μl kinase buffer (containing10% glycerol, 1 mM DTT, 10 mM MnCl₂, 20 nM microcystin, proteaseinhibitors, 1 μg of the indicated substrate, 10 μM ATP, and 10 μCi[γ-32P]ATP (6000 Ci/mmole) (NEN)] was added to each sample, and kinasereactions were performed for 30 min at 30° C. Reactions were terminatedby addition of 40 μl of 4×-SDS-PAGE sample buffer, and heating to 100°C.

EXAMPLE 4 Subcellular Localization of ATX

[0125] The subcellular localization of ATX was examined by biochemicalfractionation of U2OS cells, followed by immunoprecipitation of crudenuclear and cytoplasmic fractions with α-ATX Ab-2. Comparable levels ofATX were found in the nuclear and cytoplasmic extracts from U2OS cells(data not shown). The integrity of these subcellular fractions wasconfirmed by immunoprecipitation and immunoblotting of parallel sampleswith antibodies specific for PLCγ1 and ATR, which are localized to thecytoplasm and nucleus, respectively. The presence of ATX in both thecytoplasmic and nuclear compartments was further documented byimmunostaining of U2OS cells with affinity-purified α-ATX Ab-2. Exposureof cells to genotoxic agents-triggers the appearance of DNAdamage-induced nuclear foci containing either ATM or ATR (Andegeko etal., J. Biol. Chem. 276:38334-38230 (2001); Tibbetts et al., Genes andDevelopment 14:2989-3002 (2000)). To determine whether ATX undergoessimilar changes in subcellular localization in response to genotoxicstress, U2OS cells were treated with 400 J/m2 UV-B, and stained withα-ATX Ab-2. Exposure to UV triggered the appearance of ATX-containingnuclear foci. The ATX-positive foci were evident within 1 h after UVtreatment, and continued to accumulate in the cells until at least 8 hpost-treatment, at which time greater than 50% of the cells exhibitedmultiple ATX-containing foci. In contrast, the formation of ATX fociafter treatment of U2OS cells with 20 Gy IR was not detected.

[0126] In addition, the effect of genotoxic stress on the protein kinaseactivity of ATX in immune complex assays was determined. Consistent withthe results of the immunofluorescence staining experiments, treatment ofthe cells with IR failed to induce a reproducible increase in theprotein kinase activity observed in α-ATX immunoprecipitates (data notshown). On the other hand, UV exposure caused a modest but consistentincrease in ATX kinase activity that reached a maximal level at 4 hpost-irradiation. Collectively, the results of the nuclear localizationand protein kinase activity studies indicated that, like ATM and ATR,ATX participated in cellular responses to DNA damage or other forms ofstress induced by UV irradiation.

[0127] Methods:

[0128] Cell Fractionation, Immunoprecipitation, and Immunoblotting

[0129] For subcellular fractionations, U2OS cells were resuspended incold homogenization buffer (25 mM Hepes, pH 7.4, 250 mM sucrose, 1 mMEGTA, 5 mM MgCl2, 50 mM NaF, 1 mM DTT, plus protease inhibitors) andDounce homogenized on ice with 40 strokes in a Tefloncoated homogenizer.The nuclei were pelleted at 500×g, and the supernatant was collected asthe crude cytoplasmic fraction. Prior to immunoprecipitation, 150 mMNaCl and 1% (wt/vol) NP-40 (final concentration) were added to the crudecytoplasmic fractions. Nuclear extracts were prepared by suspending thenuclear pellets in extraction buffer (50 mM Tris-HCl, pH 7.4, 150 mMNaCl, 1% NP-40, 1 mM dithiothreitol), supplemented with proteaseinhibitors (10 μg per ml leupeptin, 10 μg per ml aprotinin, 1 μMpepstatin). After 15 min on ice, the samples were centrifuged, and thesupernatant was collected for analysis. For immunoprecipitations, cellextracts were prepared by resuspending washed cell pellets in lysisbuffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% NP-40, 1 mMdithiothreitol) plus protease inhibitors. When samples were prepared forimmune complex kinase assays, the lysis buffer was modified to contain1% Tween-20 detergent in place of NP-40, and additional phosphataseinhibitors (20 mM β-glycerol phosphate and 50 nM microcystin). Sampleswere incubated on ice for 15 min, and then clarified by centrifugation.HA-tagged recombinant proteins were immunoprecipitated from cellextracts with 4 μg of α-HA antibody. Endogenous ATX protein wasimmunoprecipitated with 8 μg of α-ATX Ab-2. After separation bySDS-PAGE, the proteins were detected by autoradiography (for kinasereaction products) or by immunoblotting. Proteins immunoblotted withrabbit and mouse antibodies were detected with protein A-horseradishperoxidase (HRP) (Amersham), and sheep anti-mouse IgG-HRP (Amersham),respectively. Immunoreactive proteins were illuminated with Renaissancechemiluminescence system (NEN).

EXAMPLE 5 Effect of Decreased ATX Function on Cellular Sensitivity to UVand IR

[0130] In order to gain further insights into the role of ATX in stressresponses, U2OS cells were transfected with the kinase-inactive ATXKImutant, and the UV- and IR-sensitivities of the transfected cells inclonogenic survival assays was examined. Control cell populations weretransfected with either empty plasmid (pcDNA3.1) or with ATXWT-encodingplasmid. At 48 h post-transfection, the cells were treated with variousdoses of UV-B (FIG. 2A) or IR (FIG. 2B). The treated cells were thencultured in G418-containing medium in order to select for stablytransfected cells. Cellular survival was determined after 10 days inculture by staining emergent colonies with Coomassie blue, followed bycolorimetric quantitation of the amount of dye-bound protein present ineach sample. Expression of ATXKI, but not ATXWT, reduced the basalsurvival of otherwise untreated U2OS cells (FIG. 2B, top panel). Theseresults indicate that endogenous ATX function is required for themaintenance of normal cell viability or growth in culture. Furthermore,expression of ATXKI dramatically increased the sensitivity of U2OS cellsto the cytostatic/cytotoxic effects of both UV and IR in theseclonogenic survival assays.

[0131] To address concerns related to potential non-specific effects ofATXKI expression on cellular functions, an antisenseoligonucleotide-based approach to reduce the level of ATX expression inU2OS cells was developed. The cells were mock transfected, ortransfected with sense (S) or antisense (AS) oligonucleotides, and thentreated for 24 h with various genotoxic agents. The cells were thentrypsinized and replated, and clonogenic survival was analyzed after 10days in culture. The results obtained with AS-treated cells werestrikingly similar to those obtained with the ATXKI-expressing cells(FIGS. 2A-C). While transfection of U2OS cells with the Soligonucleotide reduced the basal level of colony outgrowth by 25%,treatment with the AS oligonucleotide decreased basal clonogenicactivity by 75% (FIG. 2C, right panel). Hence, the AS-induced decreasein ATX protein expression was accompanied by a reduction in cellviability or proliferation in the absence of environmental stress.Furthermore, the AS-treated cells were significantly more sensitive tothe suppressive effects of IR (FIG. 2C, left panel) and UV treatments(data not shown) on clonogenic survival. The reduction in ATX proteincaused by AS treatment ranged from 50-90% in over 10 independent trials.

[0132] Methods:

[0133] Cell Transfections

[0134] To prepare recombinant HA-tagged ATX and ATM proteins, HEK 293Tcells were plated onto 60-mm dishes (9×105 cells per dish), and weretransfected with 5 μg pcDNA3.1 (empty vector), HA-ATXWT, HA-ATXKI, orHA-ATMWT plasmid DNAs. Transfections were performed with the Fugene 6transfection reagent (Roche), according to the manufacturer'sinstructions. For NMD assays (see Example 9, below), U2OS cells wereseeded onto 100 mm dishes (1×106 cells per dish). After 20 h, cells weretransfected with 1.5 μg of pmCMV-G1 test plasmid, either Norm or 39Ter(Ishigaki et al., 2001); 1.5 μg of phCMV-MUP reference plasmid (Ishigakiet al., Cell 106:607-617 (2001)); and 7 μg of empty plasmid (pcDNA 3.1),plasmids encoding HA-ATMWT, HA-ATMKI, HA-ATXWT or HA-ATXKI. Forantisense transfection experiments, U2OS cells were seeded onto 60 mmdishes (1×10⁵ cells per dish) in complete medium supplemented withpenicillin/streptomycin. After 30 h, cells were either mock transfectedor transfected with sense (S) or antisense (AS) phosphorothioateoligonucleotides (Genset Oligos, La Jolla, Calif.). The Soligonucleotide spans ATX nucleotides 210-229(5′-GAGACAGGAGGGAGCTTGCT-3′), and the AS oligonucleotide iscomplementary to this sequence (5′-AGCAAGCTCCCTCCTGTCTC-3′). The cellswere transfected with oligonucleotides at final concentrations of 8μg/ml, with Fugene 6:DNA ratio of 1.6:1. Forty-eight hours aftertransfection, dishes were exposed to IR, UV-B, or 5-FU, and thenharvested for immunoblotting, cell-cycle distribution, or cell survivalassays. To examine ATX protein levels in oligonucleotide-treated cells,whole cell extracts were resolved by SDS-PAGE and immunoblotted withα-ATX Ab-1. When oligonucleotide-transfected cells were used for NMDassays, U2OS cells were seeded in culture dishes as described above. Thecells were transfected using the Fugene reagent, with 1.5 μg of pmCMV-G1test plasmid, 0.7 μg of phCMV-MUP reference plasmid, and 24 μg of S orAS oligonucleotide.

[0135] Clonogenic and G418 Survival Assays

[0136] U2OS cells were seeded into 60 mm dishes (1×105 cells per dish)in complete medium. After 48 h, cells were transfected as describedabove. Forty-eight hours after transfection, dishes were exposed to IRor UV-B, and G418 was added at 1 mg per ml in complete medium.G418-resistant cells were stained 10 days later with Coomassie Blue. Toquantitate the outgrowth of drug-resistant cells, the CoomassieBlue-bound protein was solubilized at 37

C with 0.1 M NaOH, and the soluble material was analyzed by absorbancespectroscopy at a wavelength of 590 nm. For AT4BI cell survival assays,cells were transfected with pcDNA3.1, pcDNA3.1-FLAG-ATM, or HA-ATX.After 48 h, the transfected cells were exposed to the indicated doses ofIR, and G418 was added at 8 hours post-irradiation. Drug-resistantcolonies were stained with Coomassie Blue after 10 days in culture, andthe samples were analyzed with Image Pro Plus software to quantitatecell density. For clonogenic assays where oligonucleotides were used,the cells were plated and transfected with S or AS oligonucleotides asdescribed above. Forty-eight hours after transfection, cells wereexposed to IR or UV. Twenty-four hours after exposure to damagingagents, cells were replated at 1000 cells per 60 mm dish and coloniesallowed to form for 10 days. Dishes were stained with Coomassie Blue,and the number of colonies (minimum size, 50 cells per colony) wascounted by visual examination.

EXAMPLE 6 Role of ATX in p53 Activation

[0137] A major mediator of stress-induced signaling in mammalian cellsis the tumor suppressor protein, p53 (Ko and Prives, Genes andDevelopment 10:1054-1072, (1996); Ryan et al., Curr. Opin. Cell Biol.13:332-337 (2001)). ATX phosphorylates p53 on Ser-15 (FIG. 1C), a siteimplicated in the regulation of p53 stability and transcriptionalactivity (Dumaz and Meek, Curr. Opin. Cell Biol. 13:225-231 (1999);Zhang and Xiong, Science 292:1910-1915 (2001)). Therefore, thepossibility that these two proteins are functionally linked duringcellular stress responses was investigated. U2OS cells were transientlytransfected with a HA-ATXWT or HA-ATXKI expression plasmid, togetherwith a GFP-encoding plasmid to allow for FACS-based enrichment of thetransfected cells. The GFP+ cells were then examined for IR-inducedstabilization of p53, as well as for specific phosphorylation of thisprotein on Ser-15. Expression of ATXKI strongly suppressed both thephosphorylation of Ser-15 and the overall accumulation of p53 inIR-treated cells (FIG. 3A). In contrast, overexpression of ATXWTenhanced both of these responses in cells exposed to IR. Consistent withfindings presented above, treatment of U2OS cells with the ASoligonucleotide led to a decrease in endogenous ATX expression, andconcomitantly reduced both the phosphorylation and stabilization of p53induced by IR exposure (FIG. 3B). These results indicated that ATXexhibits functional overlap with ATM during IR-induced p53 activation.

[0138] Recent findings point toward ATM as a critical upstream regulatorof the activity of the checkpoint kinase, hChk2, in IR-damaged cells(Ahn et al., Cancer Res. 60:5934-5936 (2000); Melchionna et al., Nat.Cell Biol. 2:762-765 (2000)). To determine whether ATX was also involvedin hChk2 activation, the effect of AS oligonucleotide treatment on theIR-dependent phosphorylation of hChk2 was examined. In contrast to thep53 results, the AS-treated cells retained the ability to phosphorylatehChk2 in response to IR-induced stress (FIG. 3B). These results indicatethat, while ATM and ATX serve as positive regulators of p53 function,ATX plays no identifiable role as an upstream activator of a distinctATM target protein, the hChk2 kinase. Moreover, the differential effectsof AS treatment on p53 expression versus hChk2 activation argue againstthe possibility that AS exposure leads to nonspecific inhibition ofcheckpoint signaling responses to IR-induced DNA damage.

[0139] Additional studies with AS oligonucleotide-treated cellsdemonstrated that, in contrast to ATM (Canman et al., Science 281,1677-1679. 1998; Siliciano et al., Genes Dev 11, 3471-3481 (1997)), ATXplays a role in the phosphorylation and stabilization of p53 in cellsexposed to UV light (FIG. 3C). As observed with IR as thestress-inducing agent, a reduction in ATX protein expression severelyimpaired both Ser-15 phosphorylation and p53 protein accumulation inUV-damaged cells. The recognition of UV light-induced DNA damage occursprimarily during S phase, when pyrimidine dimers and other bulky lesionsinterfere with replication fork progression (Friedberg, DNA Repair andMutagenesis (Washington, D.C., ASM Press) (1995); Lindahl and Wood, Cell103:1121-1131 (1999)).

[0140] In order to further define the potential linkage between DNAreplicational stress and ATX, the response of AS-treated cells to5-fluorouracil (5-FU), an S-phase specific cytotoxic agent was examined(Danenberg et al., Seminars in Oncology 26:621-631 (1999); Grem,Investigational New Drugs 18:299-313 (2000)). Previous findingsindicated that the cytotoxic effects of 5-FU are strongly p53-dependent(Bunz et al., J. Clin. Invest. 104:263-269 (1999)). Treatment of U2OScells with 5-FU increased p53 expression to levels similar to thoseobserved in UV-irradiated cells (FIG. 3C). However, the accumulation ofp53 induced by 5-FU exposure was not accompanied by an increase inSer-15 phosphorylation. These findings indicate that the mechanism ofp53 stabilization triggered by 5-FU does not involve upstream proteinkinases that modify the Ser-15 site. Consistent with this conclusion,the level of p53 induction by 5-FU in AS-treated cells was identical tothat observed in their S-treated counterparts. These results indicatethat the inhibitory effect of the AS treatment on p53 activation isselective for those forms of stress that induce the phosphorylation ofp53 at Ser-15.

[0141] Changes in phosphorylation at Ser-15 are typically accompanied byalterations in the expression of the p53 protein, which complicates theinterpretation of results obtained by immunoblotting of whole cellextracts with phospho-Ser-15-specific antibodies. In order to confirmthat reduced ATX expression interferes with stress-induced Ser-15phosphorylation, U2OS cells were transfected with either the S or ASoligonucleotide, and then were pretreated with the proteasome inhibitor,LLnV, to stabilize p53. In the presence of LLnV, the p53 level in eachtest population was relatively unaffected by UV exposure (data notshown). However, the ratio of phospho-Ser-15 to total p53 protein wasincreased by UV irradiation of both the mock-transfected and Soligonucleotide-treated cells. Although AS treatment partiallyinterfered with the accumulation of p53 under these conditions, thereduction in ATX expression effectively blocked the stoichiometricincrease in Ser-15 phosphorylation triggered by UV-induced stress.

EXAMPLE 7 Role of ATX in IR-induced Cell Cycle Arrest

[0142] Since p53 plays a central role in activation of the G1checkpoint, and influences S, G2, and M checkpoints as well (Giaccia andKastan, Genes & Development 12:2973-2983 (1998); Ko and Prives, supra,1996), a functional deficiency of ATX might alter the cell-cycle arrestresponses to IR and other genotoxic agents. To test this possibility,U2OS cells were pre-treated with S or AS oligonucleotides, exposed toIR, and cell-cycle distributions at 24 h post-irradiation weredetermined. In the absence of IR, AS treatment led to a reduction in thepercentage of G1 phase cells, and a concomitant accumulation of G2/Mphase cells, when compared to their S oligonucleotide-treatedcounterparts (FIG. 3D). The AS-treated cells also contained an increasedsubpopulation with <2N DNA content, which is indicative of apoptoticcells. After IR exposure, cells treated with the S oligonucleotideaccumulated in both G1 and G2/M phases and were cleared out of S phase,a profile typical of p53-positive cells that retain G1 checkpointfunction. In contrast, the AS-treated cells arrested primarily in G2/Mphase after IR exposure. The cell-cycle distribution of the AS-treatedcells was reminiscent of that observed in cells that have lostp53-dependent checkpoint function (North and Hainaut, Pathol. Biol.48:255-270 (2000); Waldman et al., Cancer Res. 55:5187-5190 (1995)).Immunoblot analyses of the same cell populations confirmed that ASexposure led to a profound reduction in ATX protein levels in U2OScells. In contrast, AS exposure caused no significant change in theexpression levels of two control proteins, ATM and PLC-γ1.

[0143] The cell-cycle distribution results described in FIG. 3Ddemonstrated that ATX-deficient cells accumulate with 4N DNA contentunder both basal culture conditions and after IR-induced stress. Thisarrest state could reflect the activation of either a G2 or a mitoticcheckpoint (or both checkpoints). To distinguish between thesepossibilities, the effects of caffeine, a known inhibitor of the G2checkpoint (Powell et al., Cancer Res. 55:1643-1648 (1995); Russell etal., Cancer Res. 55:1639-1642 (1995); Yao et al., Nat, Med. 2:1140-1143(1996)), on the cell cycle distribution of the AS-treated cells wereexamined. The G2 checkpoint inhibitor was added to the culture medium at8 h prior to harvest for determination of cell-cycle distributions (FIG.3E, left panel), and immunoblotting for ATX expression (right panel).Caffeine completely reversed the accumulation of G2/M phase cellsinduced by AS treatment, indicating that ATX deficiency triggered theactivation of a caffeine-sensitive G2 checkpoint. The immunoblottingresults confirmed that AS-treated cells displayed a marked, specificreduction in ATX protein expression. In addition, treatment of the AScells with caffeine also resulted in an increase in the percentage ofhypodiploid cells, which indicates that an intact G2 checkpointpartially protects the ATX-deficient cells from apoptotic death, forexample, by preventing a catastrophic entry into M phase.

EXAMPLE 8 ATX Overexpression Complements IR Sensitivity in ATM-DeficientCells

[0144] Based on the finding that ATM and ATX display overlappingfunctions as activators of p53, it was investigated whether ATXoverexpression can complement the phenotypic defects found in cells fromA-T patients. One characteristic defect of cells from A-T patients isreduced clonogenic survival in culture, even in the absence of DNAdsb-inducing agents (Rotman and Shiloh, Oncogene 18:6135-6144 (1998)).As shown in FIG. 4A, transient transfection of ATM-null AT4BI cells withan ATM expression plasmid increased the outgrowth of G418-resistantcolonies by approximately 2-fold, relative to cells transfected withempty vector. The clonogenic defect of AT4BI cells was partially rescued(approximately 1.5-fold increase in colony survival) by transientexpression of ATX. Thus, ATX overexpression partially complements theintrinsic clonogenic survival defect of ATM-null cells. Furthermore,low-dose (1 Gy) IR treatment sharply reduced the clonogenic 'survival ofmock-transfected AT4BI cells, and this radiosensitive phenotype wasrescued to equivalent degrees by transfection of the cells with ATM orATX (FIG. 4B). Thus, overexpression of ATX complements, at least inpart, the stress response defects observed in cells from A-T patients.

EXAMPLE,9 Roles of ATX and ATM in hUpf1-phosphorylation and NMD

[0145] The Upf1 helicase undergoes serum-inducible phosphorylation inintact cells, as demonstrated by two-dimensional(2-D) gelelectrophoresis (Pal et al., supra, 2001). Based on the evidencedisclosed herein that ATX is a UV-responsive kinase, the possibilitythat UV light exposure triggers the phosphorylation of hUpf1 inATXWT-transfected U2OS cells was investigated. Serum stimulation or UVtreatment induced virtually identical shifts in the 2-D electrophoreticmobility of hUpf1, which indicates that these agents provoke thephosphorylation of this protein at similar sites (data not shown). Incontrast, expression of the catalytically-inactive ATXKI protein blockedthe appearance of the most highly shifted form of hUpf1, and caused theaccumulation of a broad band with intermediate electrophoretic mobility.The latter hUpf1 species can be less phosphorylated forms of theprotein. Thus, overexpression of ATXKI interferes with both the serum-and UV-induced phosphorylation of hUpf1; however, these results alsoindicate that the hUpf1 is targeted by at least one additional proteinkinase in these cells.

[0146] Based on the functional overlap between ATM and ATX duringstress-induced p53 activation, it was possible that these PIKKs mightalso share the ability to regulate the RNA surveillance pathway leadingto NMD. To focus our studies of hUpf1 phosphorylation on theSer-Gln-rich region, a mammalian expression vector encoding GAL4 fusedto the carboxyl terminus (amino acids 1019-1118) of hUpf1(GAL4-hUpf11019-1118) was generated. Expression of GAL4-hUpf11019-1118in U2OS cells generates a major immunoreactive band that migrates with amolecular mass of ˜35 kDa in serum-starved cells. Stimulation of thetransfected cells with 10% fetal bovine serum or UV light leads to theincreased expression of forms of GAL4-hUpf11019-1118 that displayreduced electrophoretic mobility (FIG. 5A, left panel). In thisexperiment, cells were harvested at 6 h after serum or UV exposure;however, the GAL4-hUpf11019-1118 mobility shifts could be detected asearly as 2 h after cellular stimulation with either agent. Theappearance of these shifted GAL4-hUpf11019-1118 bands is due tophosphorylation, as treatment of the cell extracts with λ phosphatasecollapses the complex pattern of α-Gal4-reactive species into the major˜35 kDa band, which represents non-phosphorylated GAL4-hUpf11019-1118(FIG. 5A, right panel). Moreover, the observed GAL4-hUpf11019-1118mobility shifts were due to phosphorylation of the hUpf1 fragment, asthe electrophoretic mobility GAL4 alone was not altered by cellularexposure to serum or UV (FIG. 5A, lower panel). In the experiment shownin FIG. 5A (left panel), selected samples were pretreated with 20 μMwortmannin in order to inhibit endogenous ATX and ATM kinase activities(Sarkaria et al., supra, 1998)). In the wortmannin-treated cells, theFBS- or UV-induced generation of the most slowly migrating form ofGAL4-hUpf11019-1118 (indicated with an arrow) was preferentiallyinhibited. This drug effect was accompanied by an increase in theabundance of the less shifted bands, which can represent lessphosphorylated forms of GAL4-hUpf11019-1118., These results indicatethat, although a wortmannin-sensitive protein kinase(s) contributes tothe inducible phosphorylation of the GAL4-hUpf11019-1118 reporterprotein, the hUpf1 carboxyl-terminal region is also targeted formodification by at least one additional, wortmannin-resistant proteinkinase.

[0147] To further examine the contributions of ATM and ATX to thephosphorylation of GAL4-hUpf11019-1118, U2OS cells were cotransfectedwith wild-type (WT) or kinase-inactive (KI) versions of HA-ATM orHA-ATX. Expression of either HA-ATMKI or HA-ATXKI strongly suppressedthe phosphorylation of GAL4-hUpf11019-1118 in cells treated with UVlight (FIG. 5B) or serum (data not shown). Expression of thecatalytically active HA-ATMWT or HA-ATXWT proteins enhanced thephosphorylation of GAL4-hUpf11019-1118 in both unstimulated andstimulated cells. The latter results add further support to the notionthat ATX and ATM are capable of phosphorylating hUpf1 carboxyl-terminalregion in intact cells. In these experiments, the HA-tagged ATM and ATXproteins were overexpressed by approximately 2- and 1.5-fold,respectively, when compared to their endogenous counterparts (data notshown).

[0148] In addition, the effects of HA-ATXKI and HA-ATMKI overexpressionon NMD were comparatively examined using an established assay (Sun etal., Proc. Natl. Acad. Sci. USA 95:10009-1998). U2OS cells weretransfected with a plasmid encoding either the normal human β-globingene (Norm) or a mutated β-globin gene bearing a premature terminationcodon (Ter), together with a reference plasmid encoding the mouseurinary protein (MUP). Where indicated, the cells were co-transfectedwith empty vector, or expression vectors encoding wild-type orkinase-inactive versions of ATM (HA-ATMWT, HA-ATMKI) or ATX (HA-ATXWT,HA-ATXKI) (FIG. 6A). Expression of the kinase-inactive HA-ATMKI orHA-ATXKI proteins abrogated NMD of the Ter-containing β-globin mRNA.Furthermore, treatment of the cells with the AS oligonucleotide toreduce endogenous ATX expression confirmed that ATX expression isrequired for maximal NMD activity under these assay conditions (FIG.6B). Collectively, these results indicate that ATM and ATX function asshared components of the pathways leading to both NMD and p53 activationduring UV- and IR-induced stress.

[0149] Methods:

[0150] Construction of GST and GAL4-hUpf1 Fusion Proteins

[0151] The hUpf11019-1118-BamHI fragment was generated by PCRamplification of full-length hUpf1 using the following primers:5′-AGGAGGGGATCCGGACGCCAGAAGAACCGCTTTGGG-3′,5′-AGGAGGGGATCCATACTGGGACAGCCCCGTCAC-3′. This fragment was subclonedinto the BamHI site of pGEX-2T and pCMX-GAL4(N) to generate theGSThUpf11019-1118 and GAL4-hUpf11019-1118 fusion proteins, respectively.

[0152] GAL4-hUpf1 Mobility Shift Assays

[0153] U2OS cells were plated in 60 mm dishes (4×105 cells per dish),and then transfected with 0.5 μg pCMX-GAL4 or pCMX-GAL4-hUpf11019-1118,together with 4.5 μg pcDNA3.1-HA-ATXWT, HA-ATXKI, HA-ATMWT, or HA-ATMKIplasmid DNAs. The HA-ATMKI protein contains an Asp-2870>Ala mutationthat inactivates the kinase domain. Twenty hours after transfection,serum was removed from the medium, and the cells were cultured for anadditional 24 h. The cells were then treated with 10% fetal bovine serumor 100 J/m2 UV-B. Where indicated, the serum-starved cells werepretreated for 30 min with 20 μM wortmannin prior to treatment withserum or UV. Cells were harvested in lysis buffer containing 25 mMHepes, pH 7.4, 300 mM NaCl, 1.5 mM MgCl2, 1 mM EGTA, 1% Triton X-100, 20mM β-glycerophosphate, 20 nM microcystin, 0.1 mM sodium orthovanadate, 1mM DTT, plus protease inhibitors. For phosphatase treatment, 600 U λphosphatase was added to cellular extracts (New England Biolabs). Cellextracts were resolved on by SDS-PAGE, and were immunoblotted withα-GAL4 antibody.

[0154] RNA Isolation and Assays of NMD

[0155] Total or nuclear RNA was isolated using Trizol (Invitrogen) orthe NE-PER kit (Pierce), respectively. The extent of NMD was determinedby using RT-PCR to quantitate the levels of Globin and MUP mRNA asdescribed previously (Ishigaki et al., Cell 83: 1-4 (2001)), except that21 cycles of PCR were used when analyzing the effects of ATX-specific Sand AS oligonucleotides.

EXAMPLE 10 Decreased Expression of ATX Results in SpontaneouslyIncreased DNA Damage

[0156] U2OS osteosarcoma cells were treated with small-interfering (si)RNA directed against luciferase (control) or ATX. The siRNA consistentlyinduces a >80% decrease in the expression of ATX protein at day 3post-transfection. The transfected cells were treated with 0, 3, or 10Gy ionizing radiation (IR), and then lysed after either 1 hour or 18hours. The cell extracts were separated by SDS-PAGE and immunoblottedwith an anti-phospho-Histone H2AX antibody. A significant increase inphospho-Histone H2AX immunoreactivity in 0 Gy IR-treated cells exposedto the ATX siRNA relative to the corresponding luciferasesiRNA-transfected control cells was detected (see FIG. 7). The level ofa control polypeptide, α-tubulin, was also determined to show equalloading of each lane.

[0157] The appearance of phospho-Histone H2AX marks DNA double-strandbreaks or other forms of DNA damage, as indicated by the stronginduction of immunoreactivity in IR-treated cells. These resultsindicate that ATX-deficient cells can spontaneously develop DNA damage.Furthermore, the compromised phosphorylation of Histone H2AX observed inIR-treated, ATX-deficient cells indicates that loss of ATX function canalso compromise DNA damage recognition and/or repair in mammalian cells.

[0158] Throughout this application various publications have beenreferenced within parentheses. The disclosures of these publications intheir entireties are hereby incorporated by reference in thisapplication in order to more fully describe the state of the art towhich this invention pertains.

[0159] Although the invention has been described with reference to thedisclosed embodiments, those skilled in the art will readily appreciatethat the specific experiments detailed are only illustrative of theinvention. It should be understood that various modifications can bemade without departing from the spirit of the invention.

1 11 1 12464 DNA Homo sapiens CDS (91)...(10656) 1 caaccttcaa attcagctgtggtgtctcgg caaaggcacg atgataccag agtccacgct 60 gacatacaga atgacgaaaaggagagatcg atg tct tat tgt gat gag tct cga 114 Met Ser Tyr Cys Asp GluSer Arg 1 5 ctg tcg aat ctt ctt cgg agg atc acc cgg gaa gac gac aga gaccga 162 Leu Ser Asn Leu Leu Arg Arg Ile Thr Arg Glu Asp Asp Arg Asp Arg10 15 20 aga ttg gct act gta aag cag ttg aaa gaa ttt att cag caa cca gaa210 Arg Leu Ala Thr Val Lys Gln Leu Lys Glu Phe Ile Gln Gln Pro Glu 2530 35 40 aat aag ctg gta cta gtt aaa caa ttg gat aat atc ttg gct gct gta258 Asn Lys Leu Val Leu Val Lys Gln Leu Asp Asn Ile Leu Ala Ala Val 4550 55 cat gac gtg ctt aat gaa agt agc aaa ttg ctt cag gag ttg aga cag306 His Asp Val Leu Asn Glu Ser Ser Lys Leu Leu Gln Glu Leu Arg Gln 6065 70 gag gga gct tgc tgt ctt ggc ctt ctt tgt gct tct ctg agc tat gag354 Glu Gly Ala Cys Cys Leu Gly Leu Leu Cys Ala Ser Leu Ser Tyr Glu 7580 85 gct gag aag atc ttc aag tgg att ttt agc aaa ttt agc tca tct gca402 Ala Glu Lys Ile Phe Lys Trp Ile Phe Ser Lys Phe Ser Ser Ser Ala 9095 100 aaa gat gaa gtt aaa ctc ctc tac tta tgt gcc acc tac aaa gca cta450 Lys Asp Glu Val Lys Leu Leu Tyr Leu Cys Ala Thr Tyr Lys Ala Leu 105110 115 120 gag act gta gga gaa aag aaa gcc ttt tca tct gta atg cag cttgta 498 Glu Thr Val Gly Glu Lys Lys Ala Phe Ser Ser Val Met Gln Leu Val125 130 135 atg acc agc ctg cag tct att ctt gaa aat gtg gat aca cca gaattg 546 Met Thr Ser Leu Gln Ser Ile Leu Glu Asn Val Asp Thr Pro Glu Leu140 145 150 ctt tgt aaa tgt gtt aag tgc att ctt ttg gtg gct cga tgt taccct 594 Leu Cys Lys Cys Val Lys Cys Ile Leu Leu Val Ala Arg Cys Tyr Pro155 160 165 cat att ttc agc act aat ttt agg gat aca gtt gat ata tta gttgga 642 His Ile Phe Ser Thr Asn Phe Arg Asp Thr Val Asp Ile Leu Val Gly170 175 180 tgg cat ata gat cat act cag aaa cct tcg ctc acg cag cag gtatct 690 Trp His Ile Asp His Thr Gln Lys Pro Ser Leu Thr Gln Gln Val Ser185 190 195 200 ggg tgg ttg cag agt ttg gag cca ttt tgg gta gct gat cttgca ttt 738 Gly Trp Leu Gln Ser Leu Glu Pro Phe Trp Val Ala Asp Leu AlaPhe 205 210 215 tct act act ctt ctt ggt cag ttt ctg gaa gac atg gaa gcatat gct 786 Ser Thr Thr Leu Leu Gly Gln Phe Leu Glu Asp Met Glu Ala TyrAla 220 225 230 gag gac ctc agc cat gtg gcc tct ggg gaa tca gtg gat gaagat gtc 834 Glu Asp Leu Ser His Val Ala Ser Gly Glu Ser Val Asp Glu AspVal 235 240 245 cct cct cca tca gtg tca tta cca aag ctg gct gca ctt ctccgg gta 882 Pro Pro Pro Ser Val Ser Leu Pro Lys Leu Ala Ala Leu Leu ArgVal 250 255 260 ttt agt act gtg gtg agg agc att ggg gaa cgc ttc agc ccaatt cgg 930 Phe Ser Thr Val Val Arg Ser Ile Gly Glu Arg Phe Ser Pro IleArg 265 270 275 280 ggt cct cca att act gag gca tat gta aca gat gtt ctgtac aga gta 978 Gly Pro Pro Ile Thr Glu Ala Tyr Val Thr Asp Val Leu TyrArg Val 285 290 295 atg aga tgt gtg acg gct gca aac cag gtg ttt ttt tctgag gct gtg 1026 Met Arg Cys Val Thr Ala Ala Asn Gln Val Phe Phe Ser GluAla Val 300 305 310 ttg aca gct gct aat gag tgt gtt ggt gtt ttg ctc ggcagc ttg gat 1074 Leu Thr Ala Ala Asn Glu Cys Val Gly Val Leu Leu Gly SerLeu Asp 315 320 325 cct agc atg act ata cat tgt gac atg gtc att aca tatgga tta gac 1122 Pro Ser Met Thr Ile His Cys Asp Met Val Ile Thr Tyr GlyLeu Asp 330 335 340 caa ctg gag aat tgc cag act tgt ggt acc gat tat atcatc tca gtc 1170 Gln Leu Glu Asn Cys Gln Thr Cys Gly Thr Asp Tyr Ile IleSer Val 345 350 355 360 ttg aat tta ctc acg ctg att gtt gaa cag ata aatacg aaa ctg cca 1218 Leu Asn Leu Leu Thr Leu Ile Val Glu Gln Ile Asn ThrLys Leu Pro 365 370 375 tca tca ttt gta gaa aaa ctg ttt ata cca tca tctaaa cta cta ttc 1266 Ser Ser Phe Val Glu Lys Leu Phe Ile Pro Ser Ser LysLeu Leu Phe 380 385 390 ttg cgt tat cat aaa gaa aaa gag gtt gtt gct gtagcc cat gct gtt 1314 Leu Arg Tyr His Lys Glu Lys Glu Val Val Ala Val AlaHis Ala Val 395 400 405 tat caa gca gtg ctc agc ttg aag aat att cct gttttg gag act gcc 1362 Tyr Gln Ala Val Leu Ser Leu Lys Asn Ile Pro Val LeuGlu Thr Ala 410 415 420 tat aag tta ata ttg gga gaa atg act tgt gcc ctaaac aac ctc cta 1410 Tyr Lys Leu Ile Leu Gly Glu Met Thr Cys Ala Leu AsnAsn Leu Leu 425 430 435 440 cac agt cta caa ctt cct gag gcc tgt tct gaaata aaa cat gag gct 1458 His Ser Leu Gln Leu Pro Glu Ala Cys Ser Glu IleLys His Glu Ala 445 450 455 ttt aag aat cat gtg ttc aat gta gac aat gcaaaa ttt gta gtt ata 1506 Phe Lys Asn His Val Phe Asn Val Asp Asn Ala LysPhe Val Val Ile 460 465 470 ttt gac ctc agt gcc ctg act aca att gga aatgcc aaa aac tca cta 1554 Phe Asp Leu Ser Ala Leu Thr Thr Ile Gly Asn AlaLys Asn Ser Leu 475 480 485 ata ggg atg tgg gcg cta tct cca act gtc tttgca ctt ctg agt aag 1602 Ile Gly Met Trp Ala Leu Ser Pro Thr Val Phe AlaLeu Leu Ser Lys 490 495 500 aat ctg atg att gtg cac agt gac ctg gct gttcac ttc cct gcc att 1650 Asn Leu Met Ile Val His Ser Asp Leu Ala Val HisPhe Pro Ala Ile 505 510 515 520 cag tat gct gtg ctc tac aca ttg tat tctcat tgt acc agg cat gat 1698 Gln Tyr Ala Val Leu Tyr Thr Leu Tyr Ser HisCys Thr Arg His Asp 525 530 535 cac ttt atc tct agt agc ctc agt tct tcctct cct tct ttg ttt gat 1746 His Phe Ile Ser Ser Ser Leu Ser Ser Ser SerPro Ser Leu Phe Asp 540 545 550 gga gct gtg att agc act gta act acg gctaca aag aaa cat ttc tca 1794 Gly Ala Val Ile Ser Thr Val Thr Thr Ala ThrLys Lys His Phe Ser 555 560 565 att ata tta aat ctt ctg gga ata tta cttaag aaa gat aac ctt aac 1842 Ile Ile Leu Asn Leu Leu Gly Ile Leu Leu LysLys Asp Asn Leu Asn 570 575 580 cag gac acg agg aaa ctg tta atg act tgggct ttg gaa gca gct gtt 1890 Gln Asp Thr Arg Lys Leu Leu Met Thr Trp AlaLeu Glu Ala Ala Val 585 590 595 600 tta atg aag aag tct gaa aca tac gcacct tta ttc tct ctt ccg tct 1938 Leu Met Lys Lys Ser Glu Thr Tyr Ala ProLeu Phe Ser Leu Pro Ser 605 610 615 ttc cat aaa ttt tgc aaa ggc ctt ttagcc aac act ctc gtt gaa gat 1986 Phe His Lys Phe Cys Lys Gly Leu Leu AlaAsn Thr Leu Val Glu Asp 620 625 630 gtg aat atc tgt ctg cag gca tgc agcagt cta cat gct ctg tcc tct 2034 Val Asn Ile Cys Leu Gln Ala Cys Ser SerLeu His Ala Leu Ser Ser 635 640 645 tcc ttg cca gat gat ctt tta cag agatgt gtc gat gtt tgc cgt gtt 2082 Ser Leu Pro Asp Asp Leu Leu Gln Arg CysVal Asp Val Cys Arg Val 650 655 660 caa cta gtg cac agt gga act cgt attcga caa gca ttt gga aaa ctg 2130 Gln Leu Val His Ser Gly Thr Arg Ile ArgGln Ala Phe Gly Lys Leu 665 670 675 680 ttg aaa tca att cct tta gat gttgtc cta agc aat aac aat cac aca 2178 Leu Lys Ser Ile Pro Leu Asp Val ValLeu Ser Asn Asn Asn His Thr 685 690 695 gaa att caa gaa att tct tta gcatta aga agt cac atg agt aaa gca 2226 Glu Ile Gln Glu Ile Ser Leu Ala LeuArg Ser His Met Ser Lys Ala 700 705 710 cca agt aat aca ttc cac ccc caagat ttc tct gat gtt att agt ttt 2274 Pro Ser Asn Thr Phe His Pro Gln AspPhe Ser Asp Val Ile Ser Phe 715 720 725 att ttg tat ggg aac tct cat agaaca ggg aag gac aat tgg ttg gaa 2322 Ile Leu Tyr Gly Asn Ser His Arg ThrGly Lys Asp Asn Trp Leu Glu 730 735 740 aga ctg ttc tat agc tgc cag agactg gat aag cgt gac cag tca aca 2370 Arg Leu Phe Tyr Ser Cys Gln Arg LeuAsp Lys Arg Asp Gln Ser Thr 745 750 755 760 att cca cgc aat ctc ctg aagaca gat gct gtc ctt tgg cag tgg gcc 2418 Ile Pro Arg Asn Leu Leu Lys ThrAsp Ala Val Leu Trp Gln Trp Ala 765 770 775 ata tgg gaa gct gca caa ttcact gtt ctt tct aag ctg aga acc cca 2466 Ile Trp Glu Ala Ala Gln Phe ThrVal Leu Ser Lys Leu Arg Thr Pro 780 785 790 ctg ggc aga gct caa gac accttc cag aca att gaa ggt atc att cga 2514 Leu Gly Arg Ala Gln Asp Thr PheGln Thr Ile Glu Gly Ile Ile Arg 795 800 805 agt ctc gca gct cac aca ttaaac cct gat cag gat gtt agt cag tgg 2562 Ser Leu Ala Ala His Thr Leu AsnPro Asp Gln Asp Val Ser Gln Trp 810 815 820 aca act gca gac aat gat gaaggc cat ggt aac aac caa ctt aga ctt 2610 Thr Thr Ala Asp Asn Asp Glu GlyHis Gly Asn Asn Gln Leu Arg Leu 825 830 835 840 gtt ctt ctt ctg cag tatctg gaa aat ctg gag aaa tta atg tat aat 2658 Val Leu Leu Leu Gln Tyr LeuGlu Asn Leu Glu Lys Leu Met Tyr Asn 845 850 855 gca tac gag gga tgt gctaat gca tta act tca cct ccc aag gtc att 2706 Ala Tyr Glu Gly Cys Ala AsnAla Leu Thr Ser Pro Pro Lys Val Ile 860 865 870 aga act ttt ttc tat accaat cgc caa act tgt cag gac tgg cta acg 2754 Arg Thr Phe Phe Tyr Thr AsnArg Gln Thr Cys Gln Asp Trp Leu Thr 875 880 885 cgg att cga ctc tcc atcatg agg gta gga ttg ttg gca ggc cag cct 2802 Arg Ile Arg Leu Ser Ile MetArg Val Gly Leu Leu Ala Gly Gln Pro 890 895 900 gca gtg aca gtg aga catggc ttt gac ttg ctt aca gag atg aaa aca 2850 Ala Val Thr Val Arg His GlyPhe Asp Leu Leu Thr Glu Met Lys Thr 905 910 915 920 acc agc cta tct cagggg aat gaa ttg gaa gta acc att atg atg gtg 2898 Thr Ser Leu Ser Gln GlyAsn Glu Leu Glu Val Thr Ile Met Met Val 925 930 935 gta gaa gca tta tgtgaa ctt cat tgt cct gaa gct ata cag gga att 2946 Val Glu Ala Leu Cys GluLeu His Cys Pro Glu Ala Ile Gln Gly Ile 940 945 950 gct gtc tgg tca tcatct att gtt gga aaa aat ctt ctg tgg att aac 2994 Ala Val Trp Ser Ser SerIle Val Gly Lys Asn Leu Leu Trp Ile Asn 955 960 965 tca gtg gct caa caggct gaa ggg agg ttt gaa aag gcc tct gtg gag 3042 Ser Val Ala Gln Gln AlaGlu Gly Arg Phe Glu Lys Ala Ser Val Glu 970 975 980 tac cag gaa cac ctgtgt gcc atg aca ggt gtt gat tgc tgc atc tcc 3090 Tyr Gln Glu His Leu CysAla Met Thr Gly Val Asp Cys Cys Ile Ser 985 990 995 1000 agc ttt gac aaatcg gtg ctc acc tta gcc aat gct ggg cgt aac agt 3138 Ser Phe Asp Lys SerVal Leu Thr Leu Ala Asn Ala Gly Arg Asn Ser 1005 1010 1015 gcc agc ccgaaa cat tct ctg aat ggt gaa tcc aga aaa act gtg ctg 3186 Ala Ser Pro LysHis Ser Leu Asn Gly Glu Ser Arg Lys Thr Val Leu 1020 1025 1030 tcc aaaccg act gac tct tcc cct gag gtt ata aat tat tta gga aat 3234 Ser Lys ProThr Asp Ser Ser Pro Glu Val Ile Asn Tyr Leu Gly Asn 1035 1040 1045 aaagca tgt gag tgc tac atc tca att gcc gat tgg gct gct gtg cag 3282 Lys AlaCys Glu Cys Tyr Ile Ser Ile Ala Asp Trp Ala Ala Val Gln 1050 1055 1060gaa tgg cag aac gct atc cat gac ttg aaa aag agt acc agt agc act 3330 GluTrp Gln Asn Ala Ile His Asp Leu Lys Lys Ser Thr Ser Ser Thr 1065 10701075 1080 tcc ctc aac ctg aaa gct gac ttc aac tat ata aaa tca tta agcagc 3378 Ser Leu Asn Leu Lys Ala Asp Phe Asn Tyr Ile Lys Ser Leu Ser Ser1085 1090 1095 ttt gag tct gga aaa ttt gtt gaa tgt acc gag caa tta gaattg tta 3426 Phe Glu Ser Gly Lys Phe Val Glu Cys Thr Glu Gln Leu Glu LeuLeu 1100 1105 1110 cca gga gaa aat atc aat cta ctt gct gga gga tca aaagaa aaa ata 3474 Pro Gly Glu Asn Ile Asn Leu Leu Ala Gly Gly Ser Lys GluLys Ile 1115 1120 1125 gac atg aaa aaa ctg ctt cct aac atg tta agt ccggat ccg agg gaa 3522 Asp Met Lys Lys Leu Leu Pro Asn Met Leu Ser Pro AspPro Arg Glu 1130 1135 1140 ctt cag aaa tcc att gaa gtt caa ttg tta agaagt tct gtt tgt ttg 3570 Leu Gln Lys Ser Ile Glu Val Gln Leu Leu Arg SerSer Val Cys Leu 1145 1150 1155 1160 gca act gct tta aac ccg ata gaa caagat cag aag tgg cag tct ata 3618 Ala Thr Ala Leu Asn Pro Ile Glu Gln AspGln Lys Trp Gln Ser Ile 1165 1170 1175 act gaa aat gtg gta aag tac ttgaag caa aca tcc cgc atc gct att 3666 Thr Glu Asn Val Val Lys Tyr Leu LysGln Thr Ser Arg Ile Ala Ile 1180 1185 1190 gga cct ctg aga ctt tct acttta aca gtt tca cag tct ttg cca gtt 3714 Gly Pro Leu Arg Leu Ser Thr LeuThr Val Ser Gln Ser Leu Pro Val 1195 1200 1205 cta agt acc ttg cag ctgtat tgc tca tct gct ttg gag aac aca gtt 3762 Leu Ser Thr Leu Gln Leu TyrCys Ser Ser Ala Leu Glu Asn Thr Val 1210 1215 1220 tct aac aga ctt tcaaca gag gac tgt ctt att cca ctc ttc agt gaa 3810 Ser Asn Arg Leu Ser ThrGlu Asp Cys Leu Ile Pro Leu Phe Ser Glu 1225 1230 1235 1240 gct tta cgttca tgt aaa cag cat gac gtg agg cca tgg atg cag gca 3858 Ala Leu Arg SerCys Lys Gln His Asp Val Arg Pro Trp Met Gln Ala 1245 1250 1255 tta aggtat act atg tac cag aat cag ttg ttg gag aaa att aaa gaa 3906 Leu Arg TyrThr Met Tyr Gln Asn Gln Leu Leu Glu Lys Ile Lys Glu 1260 1265 1270 caaaca gtc cca att aga agc cat ctc atg gaa tta ggt cta aca gca 3954 Gln ThrVal Pro Ile Arg Ser His Leu Met Glu Leu Gly Leu Thr Ala 1275 1280 1285gca aaa ttt gct aga aaa cga ggg aat gtg tcc ctt gca aca aga ctg 4002 AlaLys Phe Ala Arg Lys Arg Gly Asn Val Ser Leu Ala Thr Arg Leu 1290 12951300 ctg gca cag tgc agt gaa gtt cag ctg gga aag acc acc act gca cag4050 Leu Ala Gln Cys Ser Glu Val Gln Leu Gly Lys Thr Thr Thr Ala Gln1305 1310 1315 1320 gat tta gtc caa cat ttt aaa aaa cta tca acc caa ggtcaa gtg gat 4098 Asp Leu Val Gln His Phe Lys Lys Leu Ser Thr Gln Gly GlnVal Asp 1325 1330 1335 gaa aaa tgg ggg ccc gaa ctt gat att gaa aaa accaaa ttg ctt tat 4146 Glu Lys Trp Gly Pro Glu Leu Asp Ile Glu Lys Thr LysLeu Leu Tyr 1340 1345 1350 aca gca ggc cag tca aca cat gca atg gaa atgttg agt tct tgt gcc 4194 Thr Ala Gly Gln Ser Thr His Ala Met Glu Met LeuSer Ser Cys Ala 1355 1360 1365 ata tct ttc tgc aag tct gtg aaa gct gaatat gca gtt gct aaa tca 4242 Ile Ser Phe Cys Lys Ser Val Lys Ala Glu TyrAla Val Ala Lys Ser 1370 1375 1380 att ctg aca ctg gct aaa tgg atc caggca gaa tgg aaa gag att tca 4290 Ile Leu Thr Leu Ala Lys Trp Ile Gln AlaGlu Trp Lys Glu Ile Ser 1385 1390 1395 1400 gga cag ctg aaa cag gtt tacaga gct cag cac caa cag aac ttc aca 4338 Gly Gln Leu Lys Gln Val Tyr ArgAla Gln His Gln Gln Asn Phe Thr 1405 1410 1415 ggt ctt tct act ttg tctaaa aac ata ctc act cta ata gaa ctg cca 4386 Gly Leu Ser Thr Leu Ser LysAsn Ile Leu Thr Leu Ile Glu Leu Pro 1420 1425 1430 tct gtt aat acg atggaa gaa gag tat cct cgg atc gag agt gaa tct 4434 Ser Val Asn Thr Met GluGlu Glu Tyr Pro Arg Ile Glu Ser Glu Ser 1435 1440 1445 aca gtg cat attgga gtt gga gaa cct gac ttc att ttg gga cag ttg 4482 Thr Val His Ile GlyVal Gly Glu Pro Asp Phe Ile Leu Gly Gln Leu 1450 1455 1460 tat cac ctgtct tca gta cag gca cct gaa gta gcc aaa tct tgg gca 4530 Tyr His Leu SerSer Val Gln Ala Pro Glu Val Ala Lys Ser Trp Ala 1465 1470 1475 1480 gcgttg gcc agc tgg gct tat agg tgg ggc aga aag gtg gtt gac aat 4578 Ala LeuAla Ser Trp Ala Tyr Arg Trp Gly Arg Lys Val Val Asp Asn 1485 1490 1495gcc agt cag gga gaa ggt gtt cgt ctg ctg cct aga gaa aaa tct gaa 4626 AlaSer Gln Gly Glu Gly Val Arg Leu Leu Pro Arg Glu Lys Ser Glu 1500 15051510 gtt cag aat cta ctt cca gac act ata act gag gaa gag aaa gag aga4674 Val Gln Asn Leu Leu Pro Asp Thr Ile Thr Glu Glu Glu Lys Glu Arg1515 1520 1525 ata tat ggt att ctt gga cag gct gtg tgt cgg ccg gcg gggatt cag 4722 Ile Tyr Gly Ile Leu Gly Gln Ala Val Cys Arg Pro Ala Gly IleGln 1530 1535 1540 gat gaa gat ata aca ctt cag ata act gag agt gaa gacaac gaa gaa 4770 Asp Glu Asp Ile Thr Leu Gln Ile Thr Glu Ser Glu Asp AsnGlu Glu 1545 1550 1555 1560 gat gac atg gtt gat gtt atc tgg cgt cag ttgata tca agc tgc cca 4818 Asp Asp Met Val Asp Val Ile Trp Arg Gln Leu IleSer Ser Cys Pro 1565 1570 1575 tgg ctt tca gaa ctt gat gaa agt gca actgaa gga gtt att aaa gtg 4866 Trp Leu Ser Glu Leu Asp Glu Ser Ala Thr GluGly Val Ile Lys Val 1580 1585 1590 tgg agg aaa gtt gta gat aga ata ttcagc ctg tac aaa ctc tct tgc 4914 Trp Arg Lys Val Val Asp Arg Ile Phe SerLeu Tyr Lys Leu Ser Cys 1595 1600 1605 agt gca tac ttt act ttc ctt aaactc aac gct ggt caa att cct tta 4962 Ser Ala Tyr Phe Thr Phe Leu Lys LeuAsn Ala Gly Gln Ile Pro Leu 1610 1615 1620 gat gag gat gac cct agg ctgcat tta agt cac aga gtg gaa cag agc 5010 Asp Glu Asp Asp Pro Arg Leu HisLeu Ser His Arg Val Glu Gln Ser 1625 1630 1635 1640 act gat gac atg attgtg atg gcc aca ttg cgc ctg ctg cgg ttg ctc 5058 Thr Asp Asp Met Ile ValMet Ala Thr Leu Arg Leu Leu Arg Leu Leu 1645 1650 1655 gtg aag cat gctggt gag ctt cgg cag tat ctg gag cac ggc ttg gag 5106 Val Lys His Ala GlyGlu Leu Arg Gln Tyr Leu Glu His Gly Leu Glu 1660 1665 1670 aca aca cccact gca cca tgg aga gga att att ccg caa ctt ttc tca 5154 Thr Thr Pro ThrAla Pro Trp Arg Gly Ile Ile Pro Gln Leu Phe Ser 1675 1680 1685 cgc ttaaac cac cct gaa gtg tat gtg cgc caa agt att tgt aac ctt 5202 Arg Leu AsnHis Pro Glu Val Tyr Val Arg Gln Ser Ile Cys Asn Leu 1690 1695 1700 ctctgc cgt gtg gct caa gat tcc cca cat ctc ata ttg tat cct gca 5250 Leu CysArg Val Ala Gln Asp Ser Pro His Leu Ile Leu Tyr Pro Ala 1705 1710 17151720 ata gtg ggt acc ata tcg ctt agt agt gaa tcc cag gct tca gga aat5298 Ile Val Gly Thr Ile Ser Leu Ser Ser Glu Ser Gln Ala Ser Gly Asn1725 1730 1735 aaa ttt tcc act gca att cca act tta ctt ggc aat att caagga gaa 5346 Lys Phe Ser Thr Ala Ile Pro Thr Leu Leu Gly Asn Ile Gln GlyGlu 1740 1745 1750 gaa ttg ctg gtt tct gaa tgt gag gga gga agt cct cctgca tct cag 5394 Glu Leu Leu Val Ser Glu Cys Glu Gly Gly Ser Pro Pro AlaSer Gln 1755 1760 1765 gat agc aat aag gat gaa cct aaa agt gga tta aatgaa gac caa gcc 5442 Asp Ser Asn Lys Asp Glu Pro Lys Ser Gly Leu Asn GluAsp Gln Ala 1770 1775 1780 atg atg cag gat tgt tac agc aaa att gta gataag ctg tcc tct gca 5490 Met Met Gln Asp Cys Tyr Ser Lys Ile Val Asp LysLeu Ser Ser Ala 1785 1790 1795 1800 aac ccc acc atg gta tta cag gtt cagatg ctc gtg gct gaa ctg cgc 5538 Asn Pro Thr Met Val Leu Gln Val Gln MetLeu Val Ala Glu Leu Arg 1805 1810 1815 agg gtc act gtg ctc tgg gat gagctc tgg ctg gga gtt ttg ctg caa 5586 Arg Val Thr Val Leu Trp Asp Glu LeuTrp Leu Gly Val Leu Leu Gln 1820 1825 1830 caa cac atg tat gtc ctg agacga att cag cag ctt gaa gat gag gtg 5634 Gln His Met Tyr Val Leu Arg ArgIle Gln Gln Leu Glu Asp Glu Val 1835 1840 1845 aag aga gtc cag aac aacaac acc tta cgc aaa gaa gag aaa att gca 5682 Lys Arg Val Gln Asn Asn AsnThr Leu Arg Lys Glu Glu Lys Ile Ala 1850 1855 1860 atc atg agg gag aagcac aca gct ttg atg aag ccc atc gta ttt gct 5730 Ile Met Arg Glu Lys HisThr Ala Leu Met Lys Pro Ile Val Phe Ala 1865 1870 1875 1880 ttg gag catgtg agg agt atc aca gcg gct cct gca gaa aca cct cat 5778 Leu Glu His ValArg Ser Ile Thr Ala Ala Pro Ala Glu Thr Pro His 1885 1890 1895 gaa aaatgg ttt cag gat aac tat ggt gat gcc att gaa aat gcc cta 5826 Glu Lys TrpPhe Gln Asp Asn Tyr Gly Asp Ala Ile Glu Asn Ala Leu 1900 1905 1910 gaaaaa ctg aag act cca ttg aac cct gca aag cct ggg agc agc tgg 5874 Glu LysLeu Lys Thr Pro Leu Asn Pro Ala Lys Pro Gly Ser Ser Trp 1915 1920 1925att cca ttt aaa gag ata atg cta agt ttg caa cag aga gca cag aaa 5922 IlePro Phe Lys Glu Ile Met Leu Ser Leu Gln Gln Arg Ala Gln Lys 1930 19351940 cgt gca agt tac atc ttg cgt ctt gaa gaa atc agt cca tgg ttg gct5970 Arg Ala Ser Tyr Ile Leu Arg Leu Glu Glu Ile Ser Pro Trp Leu Ala1945 1950 1955 1960 gcc atg act aac act gaa att gct ctt cct ggg gaa gtctca gcc aga 6018 Ala Met Thr Asn Thr Glu Ile Ala Leu Pro Gly Glu Val SerAla Arg 1965 1970 1975 gac act gtc aca atc cat agt gtg ggc gga acc atcaca atc tta ccg 6066 Asp Thr Val Thr Ile His Ser Val Gly Gly Thr Ile ThrIle Leu Pro 1980 1985 1990 act aaa acc aag cca aag aaa ctt ctc ttt cttgga tca gat ggg aag 6114 Thr Lys Thr Lys Pro Lys Lys Leu Leu Phe Leu GlySer Asp Gly Lys 1995 2000 2005 agc tat cct tat ctt ttc aaa gga ctg gaggat tta cat ctg gat gag 6162 Ser Tyr Pro Tyr Leu Phe Lys Gly Leu Glu AspLeu His Leu Asp Glu 2010 2015 2020 aga ata atg cag ttc cta tct att gtgaat acc atg ttt gct aca att 6210 Arg Ile Met Gln Phe Leu Ser Ile Val AsnThr Met Phe Ala Thr Ile 2025 2030 2035 2040 aat cgc caa gaa aca ccc cggttc cat gct cga cac tat tct gta aca 6258 Asn Arg Gln Glu Thr Pro Arg PheHis Ala Arg His Tyr Ser Val Thr 2045 2050 2055 cca cta gga aca aga tcagga cta atc cag tgg gta gat gga gcc aca 6306 Pro Leu Gly Thr Arg Ser GlyLeu Ile Gln Trp Val Asp Gly Ala Thr 2060 2065 2070 ccc tta ttt ggt ctttac aaa cga tgg caa caa cgg gaa gct gcc tta 6354 Pro Leu Phe Gly Leu TyrLys Arg Trp Gln Gln Arg Glu Ala Ala Leu 2075 2080 2085 caa gca caa aaggcc caa gat tcc tac caa act cct cag aat cct gga 6402 Gln Ala Gln Lys AlaGln Asp Ser Tyr Gln Thr Pro Gln Asn Pro Gly 2090 2095 2100 att gta ccccgt cct agt gaa ctt tat tac agt aaa att ggc cct gct 6450 Ile Val Pro ArgPro Ser Glu Leu Tyr Tyr Ser Lys Ile Gly Pro Ala 2105 2110 2115 2120 ttgaaa aca gtt ggg ctt agc ctg gat gtg tcc cgt cgg gat tgg cct 6498 Leu LysThr Val Gly Leu Ser Leu Asp Val Ser Arg Arg Asp Trp Pro 2125 2130 2135ctt cat gta atg aag gca gta ttg gaa gag tta atg gag gcc aca ccc 6546 LeuHis Val Met Lys Ala Val Leu Glu Glu Leu Met Glu Ala Thr Pro 2140 21452150 ccg aat ctc ctt gcc aaa gag ctc tgg tca tct tgc aca aca cct gat6594 Pro Asn Leu Leu Ala Lys Glu Leu Trp Ser Ser Cys Thr Thr Pro Asp2155 2160 2165 gaa tgg tgg aga gtt acg cag tct tat gca aga tct act gcagtc atg 6642 Glu Trp Trp Arg Val Thr Gln Ser Tyr Ala Arg Ser Thr Ala ValMet 2170 2175 2180 tct atg gtt gga tac ata att ggc ctt gga gac aga catctg gat aat 6690 Ser Met Val Gly Tyr Ile Ile Gly Leu Gly Asp Arg His LeuAsp Asn 2185 2190 2195 2200 gtt ctt ata gat atg acg act gga gaa gtt gttcac ata gat tac aat 6738 Val Leu Ile Asp Met Thr Thr Gly Glu Val Val HisIle Asp Tyr Asn 2205 2210 2215 gtt tgc ttt gaa aaa ggt aaa agc ctt agagtt cct gag aaa gta cct 6786 Val Cys Phe Glu Lys Gly Lys Ser Leu Arg ValPro Glu Lys Val Pro 2220 2225 2230 ttt cga atg aca caa aac att gaa acagca ctg ggt gta act gga gta 6834 Phe Arg Met Thr Gln Asn Ile Glu Thr AlaLeu Gly Val Thr Gly Val 2235 2240 2245 gaa ggt gta ttt agg ctt tca tgtgag cag gtt tta cac att atg cgg 6882 Glu Gly Val Phe Arg Leu Ser Cys GluGln Val Leu His Ile Met Arg 2250 2255 2260 cgt ggc aga gag acc ctg ctgacg ctg ctg gag gcc ttt gtg tac gac 6930 Arg Gly Arg Glu Thr Leu Leu ThrLeu Leu Glu Ala Phe Val Tyr Asp 2265 2270 2275 2280 cct ctg gtg gac tggaca gca gga ggc gag gct ggg ttt gct ggt gct 6978 Pro Leu Val Asp Trp ThrAla Gly Gly Glu Ala Gly Phe Ala Gly Ala 2285 2290 2295 gtc tat ggt ggaggt ggc cag cag gcc gag agc aag cag agc aag aga 7026 Val Tyr Gly Gly GlyGly Gln Gln Ala Glu Ser Lys Gln Ser Lys Arg 2300 2305 2310 gag atg gagcga gag atc acc cgc agc ctg ttt tct tct aga gta gct 7074 Glu Met Glu ArgGlu Ile Thr Arg Ser Leu Phe Ser Ser Arg Val Ala 2315 2320 2325 gag attaag gtg aac tgg ttt aag aat aga gat gag atg ctg gtt gtg 7122 Glu Ile LysVal Asn Trp Phe Lys Asn Arg Asp Glu Met Leu Val Val 2330 2335 2340 cttccc aag ttg gac ggt agc tta gat gaa tac cta agc ttg caa gag 7170 Leu ProLys Leu Asp Gly Ser Leu Asp Glu Tyr Leu Ser Leu Gln Glu 2345 2350 23552360 caa ctg aca gat gtg gaa aaa ctg cag ggc aaa cta ctg gag gaa ata7218 Gln Leu Thr Asp Val Glu Lys Leu Gln Gly Lys Leu Leu Glu Glu Ile2365 2370 2375 gag ttt cta gaa gga gct gaa ggg gtg gat cat cct tct catact ctg 7266 Glu Phe Leu Glu Gly Ala Glu Gly Val Asp His Pro Ser His ThrLeu 2380 2385 2390 caa cac agg tat tct gag cac acc caa cta cag act cagcaa aga gct 7314 Gln His Arg Tyr Ser Glu His Thr Gln Leu Gln Thr Gln GlnArg Ala 2395 2400 2405 gtt cag gaa gca atc cag gtg aag ctg aat gaa tttgaa caa tgg ata 7362 Val Gln Glu Ala Ile Gln Val Lys Leu Asn Glu Phe GluGln Trp Ile 2410 2415 2420 aca cat tat cag gct gca ttc aat aat tta gaagca aca cag ctt gca 7410 Thr His Tyr Gln Ala Ala Phe Asn Asn Leu Glu AlaThr Gln Leu Ala 2425 2430 2435 2440 agc ttg ctt caa gag ata agc aca caaatg gac ctt ggt cct cca agt 7458 Ser Leu Leu Gln Glu Ile Ser Thr Gln MetAsp Leu Gly Pro Pro Ser 2445 2450 2455 tac gtg cca gca aca gcc ttt ctgcag aat gct ggt cag gcc cac ttg 7506 Tyr Val Pro Ala Thr Ala Phe Leu GlnAsn Ala Gly Gln Ala His Leu 2460 2465 2470 att agc cag tgc gag cag ctggag ggg gag gtt ggt gct ctc ctg cag 7554 Ile Ser Gln Cys Glu Gln Leu GluGly Glu Val Gly Ala Leu Leu Gln 2475 2480 2485 cag agg cgc tcc gtg ctccgt ggc tgt ctg gag caa ctg cat cac tat 7602 Gln Arg Arg Ser Val Leu ArgGly Cys Leu Glu Gln Leu His His Tyr 2490 2495 2500 gca acc gtg gcc ctgcag tat ccg aag gcc ata ttt cag aaa cat cga 7650 Ala Thr Val Ala Leu GlnTyr Pro Lys Ala Ile Phe Gln Lys His Arg 2505 2510 2515 2520 att gaa cagtgg aag acc tgg atg gaa gag ctc atc tgt aac acc aca 7698 Ile Glu Gln TrpLys Thr Trp Met Glu Glu Leu Ile Cys Asn Thr Thr 2525 2530 2535 gta gagcgt tgt caa gag ctc tat agg aaa tat gaa atg caa tat gct 7746 Val Glu ArgCys Gln Glu Leu Tyr Arg Lys Tyr Glu Met Gln Tyr Ala 2540 2545 2550 ccccag cca ccc cca aca gtg tgt cag ttc atc act gcc act gaa atg 7794 Pro GlnPro Pro Pro Thr Val Cys Gln Phe Ile Thr Ala Thr Glu Met 2555 2560 2565acc ctg cag cga tac gca gca gac atc aac agc aga ctt att aga caa 7842 ThrLeu Gln Arg Tyr Ala Ala Asp Ile Asn Ser Arg Leu Ile Arg Gln 2570 25752580 gtg gaa cgc ttg aaa cag gaa gct gtc act gtg cca gtt tgt gaa gat7890 Val Glu Arg Leu Lys Gln Glu Ala Val Thr Val Pro Val Cys Glu Asp2585 2590 2595 2600 cag ttg aaa gaa att gaa cgt tgc att aaa gtt ttc cttcat gag aat 7938 Gln Leu Lys Glu Ile Glu Arg Cys Ile Lys Val Phe Leu HisGlu Asn 2605 2610 2615 gga gaa gaa gga tct ttg agt cta gca agt gtt attatt tct gcc ctt 7986 Gly Glu Glu Gly Ser Leu Ser Leu Ala Ser Val Ile IleSer Ala Leu 2620 2625 2630 tgt acc ctt aca agg cgt aac ctg atg atg gaaggt gca gcg tca agt 8034 Cys Thr Leu Thr Arg Arg Asn Leu Met Met Glu GlyAla Ala Ser Ser 2635 2640 2645 gct gga gaa cag ctg gtt gat ctg act tctcgg gat gga gcc tgg ttc 8082 Ala Gly Glu Gln Leu Val Asp Leu Thr Ser ArgAsp Gly Ala Trp Phe 2650 2655 2660 ttg gag gaa ctc tgc agt atg agc ggaaac gtc acc tgc ttg gtt cag 8130 Leu Glu Glu Leu Cys Ser Met Ser Gly AsnVal Thr Cys Leu Val Gln 2665 2670 2675 2680 tta ctg aag cag tgc cac ctggtg cca cag gac tta gat atc ccg aac 8178 Leu Leu Lys Gln Cys His Leu ValPro Gln Asp Leu Asp Ile Pro Asn 2685 2690 2695 ccc atg gaa gcg tct gagaca gtt cac tta gcc aat gga gtg tat acc 8226 Pro Met Glu Ala Ser Glu ThrVal His Leu Ala Asn Gly Val Tyr Thr 2700 2705 2710 tca ctt cag gaa ttgaat tcg aat ttc cgg caa atc ata ttt cca gaa 8274 Ser Leu Gln Glu Leu AsnSer Asn Phe Arg Gln Ile Ile Phe Pro Glu 2715 2720 2725 gca ctt cga tgttta atg aaa ggg gaa tac acg tta gaa agt atg ctg 8322 Ala Leu Arg Cys LeuMet Lys Gly Glu Tyr Thr Leu Glu Ser Met Leu 2730 2735 2740 cat gaa ctggac ggt ctt att gag cag acc acc gat ggc gtt ccc ctg 8370 His Glu Leu AspGly Leu Ile Glu Gln Thr Thr Asp Gly Val Pro Leu 2745 2750 2755 2760 cagact cta gtg gaa tct ctt cag gcc tac tta aga aac gca gct atg 8418 Gln ThrLeu Val Glu Ser Leu Gln Ala Tyr Leu Arg Asn Ala Ala Met 2765 2770 2775gga ctg gaa gaa gaa aca cat gct cat tac atc gat gtt gcc aga cta 8466 GlyLeu Glu Glu Glu Thr His Ala His Tyr Ile Asp Val Ala Arg Leu 2780 27852790 cta cat gct cag tac ggt gaa tta atc caa ccg aga aat ggt tca gtt8514 Leu His Ala Gln Tyr Gly Glu Leu Ile Gln Pro Arg Asn Gly Ser Val2795 2800 2805 gat gaa aca ccc aaa atg tca gct ggc cag atg ctt ttg gtagca ttc 8562 Asp Glu Thr Pro Lys Met Ser Ala Gly Gln Met Leu Leu Val AlaPhe 2810 2815 2820 gat ggc atg ttt gct caa gtt gaa act gct ttc agc ttatta gtt gaa 8610 Asp Gly Met Phe Ala Gln Val Glu Thr Ala Phe Ser Leu LeuVal Glu 2825 2830 2835 2840 aag ttg aac aag atg gaa att ccc ata gct tggcga aag att gac atc 8658 Lys Leu Asn Lys Met Glu Ile Pro Ile Ala Trp ArgLys Ile Asp Ile 2845 2850 2855 ata agg gaa gcc agg agt act caa gtt aatttt ttt gat gat gat aat 8706 Ile Arg Glu Ala Arg Ser Thr Gln Val Asn PhePhe Asp Asp Asp Asn 2860 2865 2870 cac cgg cag gtg cta gaa gag att ttcttt cta aaa aga cta cag act 8754 His Arg Gln Val Leu Glu Glu Ile Phe PheLeu Lys Arg Leu Gln Thr 2875 2880 2885 att aag gag ttc ttc agg ctc tgtggt acc ttt tct aaa aca ttg tca 8802 Ile Lys Glu Phe Phe Arg Leu Cys GlyThr Phe Ser Lys Thr Leu Ser 2890 2895 2900 gga tca agt tca ctt gaa gatcag aat act gtg aat ggg cct gta cag 8850 Gly Ser Ser Ser Leu Glu Asp GlnAsn Thr Val Asn Gly Pro Val Gln 2905 2910 2915 2920 att gtc aat gtg aaaacc ctt ttt aga aac tct tgt ttc agt gaa gac 8898 Ile Val Asn Val Lys ThrLeu Phe Arg Asn Ser Cys Phe Ser Glu Asp 2925 2930 2935 caa atg gcc aaacct atc aag gca ttc aca gct gac ttt gtg agg cag 8946 Gln Met Ala Lys ProIle Lys Ala Phe Thr Ala Asp Phe Val Arg Gln 2940 2945 2950 ctc ttg ataggg cta ccc aac caa gcc ctc gga ctc aca ctg tgc agt 8994 Leu Leu Ile GlyLeu Pro Asn Gln Ala Leu Gly Leu Thr Leu Cys Ser 2955 2960 2965 ttt atcagt gct ctg ggt gta gac atc att gct caa gta gag gca aag 9042 Phe Ile SerAla Leu Gly Val Asp Ile Ile Ala Gln Val Glu Ala Lys 2970 2975 2980 gacttt ggt gcc gaa agc aaa gtt tct gtt gat gat ctc tgt aag aaa 9090 Asp PheGly Ala Glu Ser Lys Val Ser Val Asp Asp Leu Cys Lys Lys 2985 2990 29953000 gcg gtg gaa cat aac atc cag ata ggg aag ttc tct cag ctg gtt atg9138 Ala Val Glu His Asn Ile Gln Ile Gly Lys Phe Ser Gln Leu Val Met3005 3010 3015 aac agg gca act gtg tta gca agt tct tac gac act gcc tggaag aag 9186 Asn Arg Ala Thr Val Leu Ala Ser Ser Tyr Asp Thr Ala Trp LysLys 3020 3025 3030 cat gac ttg gtg cga agg cta gaa acc agt att tct tcttgt aag aca 9234 His Asp Leu Val Arg Arg Leu Glu Thr Ser Ile Ser Ser CysLys Thr 3035 3040 3045 agc ctg cag cgg gtt cag ctg cat att gcc atg tttcag tgg caa cat 9282 Ser Leu Gln Arg Val Gln Leu His Ile Ala Met Phe GlnTrp Gln His 3050 3055 3060 gaa gat cta ctt atc aat aga cca caa gcc atgtca gtc aca cct ccc 9330 Glu Asp Leu Leu Ile Asn Arg Pro Gln Ala Met SerVal Thr Pro Pro 3065 3070 3075 3080 cca cgg tct gct atc cta acc agc atgaaa aag aag ctg cat acc ctg 9378 Pro Arg Ser Ala Ile Leu Thr Ser Met LysLys Lys Leu His Thr Leu 3085 3090 3095 agc cag att gaa act tct att gcaaca gtt cag gag aag cta gct gca 9426 Ser Gln Ile Glu Thr Ser Ile Ala ThrVal Gln Glu Lys Leu Ala Ala 3100 3105 3110 ctt gaa tca agt att gaa cagcga ctc aag tgg gca ggt ggt gcc aac 9474 Leu Glu Ser Ser Ile Glu Gln ArgLeu Lys Trp Ala Gly Gly Ala Asn 3115 3120 3125 cct gca ttg gcc cct gtacta caa gat ttt gaa gca acg ata gct gaa 9522 Pro Ala Leu Ala Pro Val LeuGln Asp Phe Glu Ala Thr Ile Ala Glu 3130 3135 3140 aga aga aat ctt gtcctt aaa gag agc caa aga gca agt cag gtc aca 9570 Arg Arg Asn Leu Val LeuLys Glu Ser Gln Arg Ala Ser Gln Val Thr 3145 3150 3155 3160 ttt ctc tgcagc aat atc att cat ttt gaa agt tta cga aca aga act 9618 Phe Leu Cys SerAsn Ile Ile His Phe Glu Ser Leu Arg Thr Arg Thr 3165 3170 3175 gca gaagcc tta aac ctg gat gcg gcg tta ttt gaa cta atc aag cga 9666 Ala Glu AlaLeu Asn Leu Asp Ala Ala Leu Phe Glu Leu Ile Lys Arg 3180 3185 3190 tgtcag cag atg tgt tcg ttt gca tca cag ttt aac agt tca gtg tct 9714 Cys GlnGln Met Cys Ser Phe Ala Ser Gln Phe Asn Ser Ser Val Ser 3195 3200 3205gag tta gag ctt cgt tta tta cag aga gtg gac act ggt ctt gaa cat 9762 GluLeu Glu Leu Arg Leu Leu Gln Arg Val Asp Thr Gly Leu Glu His 3210 32153220 cct att ggc agc tct gaa tgg ctt ttg tca gca cac aaa cag ttg acc9810 Pro Ile Gly Ser Ser Glu Trp Leu Leu Ser Ala His Lys Gln Leu Thr3225 3230 3235 3240 cag gat atg tct act cag agg gca att cag aca gag aaagag cag cag 9858 Gln Asp Met Ser Thr Gln Arg Ala Ile Gln Thr Glu Lys GluGln Gln 3245 3250 3255 ata gaa acg gtc tgt gaa aca att cag aat ctg gttgat aat ata aag 9906 Ile Glu Thr Val Cys Glu Thr Ile Gln Asn Leu Val AspAsn Ile Lys 3260 3265 3270 act gtg ctc act ggt cat aac cga cag ctt ggagat gtc aaa cat ctc 9954 Thr Val Leu Thr Gly His Asn Arg Gln Leu Gly AspVal Lys His Leu 3275 3280 3285 ttg aaa gct atg gct aag gat gaa gaa gctgct ctg gca gat ggt gaa 10002 Leu Lys Ala Met Ala Lys Asp Glu Glu AlaAla Leu Ala Asp Gly Glu 3290 3295 3300 gat gtt ccc tat gag aac agt gttagg cag ttt ttg ggt gaa tat aaa 10050 Asp Val Pro Tyr Glu Asn Ser ValArg Gln Phe Leu Gly Glu Tyr Lys 3305 3310 3315 3320 tca tgg caa gac aacatt caa aca gtt cta ttt aca tta gtc cag gct 10098 Ser Trp Gln Asp AsnIle Gln Thr Val Leu Phe Thr Leu Val Gln Ala 3325 3330 3335 atg ggt caggtt cga agt caa gaa cac gtt gaa atg ctc cag gaa atc 10146 Met Gly GlnVal Arg Ser Gln Glu His Val Glu Met Leu Gln Glu Ile 3340 3345 3350 actccc acc ttg aaa gaa ctg aaa aca caa agt cag agt atc tat aat 10194 ThrPro Thr Leu Lys Glu Leu Lys Thr Gln Ser Gln Ser Ile Tyr Asn 3355 33603365 aat tta gtg agt ttt gca tca ccc tta gtc acc gat gca aca aat gaa10242 Asn Leu Val Ser Phe Ala Ser Pro Leu Val Thr Asp Ala Thr Asn Glu3370 3375 3380 tgt tcg agt cca acg tca tct gct act tat cag cca tcc ttcgct gca 10290 Cys Ser Ser Pro Thr Ser Ser Ala Thr Tyr Gln Pro Ser PheAla Ala 3385 3390 3395 3400 gca gtc cgg agt aac act ggc cag aag act cagcct gat gtc atg tca 10338 Ala Val Arg Ser Asn Thr Gly Gln Lys Thr GlnPro Asp Val Met Ser 3405 3410 3415 cag aat gct aga aag ctg atc cag aaaaat ctt gct aca tca gct gat 10386 Gln Asn Ala Arg Lys Leu Ile Gln LysAsn Leu Ala Thr Ser Ala Asp 3420 3425 3430 act cca cca agc acc gtt ccagga act ggc aag agt gtt gct tgt agt 10434 Thr Pro Pro Ser Thr Val ProGly Thr Gly Lys Ser Val Ala Cys Ser 3435 3440 3445 cct aaa aag gca gtcaga gac cct aaa act ggg aaa gcg gtg caa gag 10482 Pro Lys Lys Ala ValArg Asp Pro Lys Thr Gly Lys Ala Val Gln Glu 3450 3455 3460 aga aac tcctat gca gtg agt gtg tgg aag aga gtg aaa gcc aag tta 10530 Arg Asn SerTyr Ala Val Ser Val Trp Lys Arg Val Lys Ala Lys Leu 3465 3470 3475 3480gag ggc cga gat gtt gat ccg aat agg agg atg tca gtt gct gaa cag 10578Glu Gly Arg Asp Val Asp Pro Asn Arg Arg Met Ser Val Ala Glu Gln 34853490 3495 gtt gac tat gtc att aag gaa gca act aat cta gat aac ttg gctcag 10626 Val Asp Tyr Val Ile Lys Glu Ala Thr Asn Leu Asp Asn Leu AlaGln 3500 3505 3510 ctg tat gaa ggt tgg aca gcc tgg gtg tga atggcaagacagtagatgag 10676 Leu Tyr Glu Gly Trp Thr Ala Trp Val * 3515 3520tctggttaag cgaggtcaga catccaccag aatcaactca gcctcaggca tccaaagcca 10736caccacagtc ggtggtgatg caactggggg cttactctga ggaaacctag gaaatctcgg 10796tgcactagga agtgaatccc gcaggacagc tgcactcagg gatacgccca acaccatggc 10856ctgcaacccc agggtcaagg gtgaaggaaa gcaagctcac cgcctgaaca cggagattgt 10916ctttctgcca cagaacagca gcagacgtgt cgggaggtta gctgcggaaa gaaatcggga 10976tgccgcggag cacagagtga tttggaactc cattccacct gaccctgtgt gtacaatcca 11036ggaaaaaaac aaaccccact cagaaacaga gaaaactggg gtcgcgaaga aatcacagcc 11096aaggaagatt tgatgcattc agattctcgt gtaacacttg ttgcttggca acagtactgg 11156ttgggttgac cagtaagtag aaaaaggcta aaggctatgc gatatgaatt tcagaaatgg 11216actgaaaatg gagagctatg taacagatac actacagtag aagaacttac ttctgaaatg 11276aagggaaaaa aaccacccca tcgttcccta ctcctcccca ccacttaccc gttccccctt 11336tacctaatct agtagattag ccatctttca aattcacttt tatttcagtc cttatatttc 11396atatacttcc gtctcgatgc tgttaacaac ttctgataac atggaaaatt caaggattgt 11456ttaaaggtct gatgatcaca cacaaaatgt aattccggtt atttaagtca tttctgtgat 11516tctatcatgt acagtttcca gaattgtcac tgtgcattca aaagtaatga atctaacaga 11576catttgattt aatgtacact cccttttgct tatagtgtgc attttttttg gaggtcattc 11636aaattttccc tcttctgtga tagctgtagt ttctttcata gaaagtagct aatccagtgt 11696aatcttttac ctttttaaaa accaagatag agtatctatt agagttttac attgttgatg 11756atagattaac aataaagtga tgttctggtg gaggtagact gaaatttttt taattcatgt 11816ttttcatttg atacttttaa tttacactta gtaaattaaa agttgtttaa tttacttggc 11876attttaggac atgtacatga aacagtgaaa atgagatcca ccaacatctt ttattaagtt 11936cagttattag tctgtgaagt gctttacttt ttgcacaatt ttaatagctt gctattcagt 11996aatacattat agtgaattca tgatcaaggt ttccttaaat ttagcattgc atttcagtac 12056tgactgtgta agctaaattg ctgatccaaa ataaaaaccc agactagaat agggttctta 12116aaatcaagta tcaatacaaa atagaacaca attaaaatct taattgttgg ctgggcacag 12176tggctcacgc ctgtaatccc agcactttgg gaggccgagg cgggcggatc atgaggttag 12236gagagcgaga ccatcctggc taacacggtg aaaccccgtc tttactaaaa tacaaaaaaa 12296attagccggg tgtggtggcg ggcgcctgta gtcccagcta ctcgggaggc tgaggcagga 12356gaatggcgtg aacccaggag gcggagcttg cagtgagccg agattgtgcc actgcactcc 12416agcctgggca acagagctag actctgtgtc aaaaataaat gactagat 12464 2 3521 PRTHomo sapiens 2 Met Ser Tyr Cys Asp Glu Ser Arg Leu Ser Asn Leu Leu ArgArg Ile 1 5 10 15 Thr Arg Glu Asp Asp Arg Asp Arg Arg Leu Ala Thr ValLys Gln Leu 20 25 30 Lys Glu Phe Ile Gln Gln Pro Glu Asn Lys Leu Val LeuVal Lys Gln 35 40 45 Leu Asp Asn Ile Leu Ala Ala Val His Asp Val Leu AsnGlu Ser Ser 50 55 60 Lys Leu Leu Gln Glu Leu Arg Gln Glu Gly Ala Cys CysLeu Gly Leu 65 70 75 80 Leu Cys Ala Ser Leu Ser Tyr Glu Ala Glu Lys IlePhe Lys Trp Ile 85 90 95 Phe Ser Lys Phe Ser Ser Ser Ala Lys Asp Glu ValLys Leu Leu Tyr 100 105 110 Leu Cys Ala Thr Tyr Lys Ala Leu Glu Thr ValGly Glu Lys Lys Ala 115 120 125 Phe Ser Ser Val Met Gln Leu Val Met ThrSer Leu Gln Ser Ile Leu 130 135 140 Glu Asn Val Asp Thr Pro Glu Leu LeuCys Lys Cys Val Lys Cys Ile 145 150 155 160 Leu Leu Val Ala Arg Cys TyrPro His Ile Phe Ser Thr Asn Phe Arg 165 170 175 Asp Thr Val Asp Ile LeuVal Gly Trp His Ile Asp His Thr Gln Lys 180 185 190 Pro Ser Leu Thr GlnGln Val Ser Gly Trp Leu Gln Ser Leu Glu Pro 195 200 205 Phe Trp Val AlaAsp Leu Ala Phe Ser Thr Thr Leu Leu Gly Gln Phe 210 215 220 Leu Glu AspMet Glu Ala Tyr Ala Glu Asp Leu Ser His Val Ala Ser 225 230 235 240 GlyGlu Ser Val Asp Glu Asp Val Pro Pro Pro Ser Val Ser Leu Pro 245 250 255Lys Leu Ala Ala Leu Leu Arg Val Phe Ser Thr Val Val Arg Ser Ile 260 265270 Gly Glu Arg Phe Ser Pro Ile Arg Gly Pro Pro Ile Thr Glu Ala Tyr 275280 285 Val Thr Asp Val Leu Tyr Arg Val Met Arg Cys Val Thr Ala Ala Asn290 295 300 Gln Val Phe Phe Ser Glu Ala Val Leu Thr Ala Ala Asn Glu CysVal 305 310 315 320 Gly Val Leu Leu Gly Ser Leu Asp Pro Ser Met Thr IleHis Cys Asp 325 330 335 Met Val Ile Thr Tyr Gly Leu Asp Gln Leu Glu AsnCys Gln Thr Cys 340 345 350 Gly Thr Asp Tyr Ile Ile Ser Val Leu Asn LeuLeu Thr Leu Ile Val 355 360 365 Glu Gln Ile Asn Thr Lys Leu Pro Ser SerPhe Val Glu Lys Leu Phe 370 375 380 Ile Pro Ser Ser Lys Leu Leu Phe LeuArg Tyr His Lys Glu Lys Glu 385 390 395 400 Val Val Ala Val Ala His AlaVal Tyr Gln Ala Val Leu Ser Leu Lys 405 410 415 Asn Ile Pro Val Leu GluThr Ala Tyr Lys Leu Ile Leu Gly Glu Met 420 425 430 Thr Cys Ala Leu AsnAsn Leu Leu His Ser Leu Gln Leu Pro Glu Ala 435 440 445 Cys Ser Glu IleLys His Glu Ala Phe Lys Asn His Val Phe Asn Val 450 455 460 Asp Asn AlaLys Phe Val Val Ile Phe Asp Leu Ser Ala Leu Thr Thr 465 470 475 480 IleGly Asn Ala Lys Asn Ser Leu Ile Gly Met Trp Ala Leu Ser Pro 485 490 495Thr Val Phe Ala Leu Leu Ser Lys Asn Leu Met Ile Val His Ser Asp 500 505510 Leu Ala Val His Phe Pro Ala Ile Gln Tyr Ala Val Leu Tyr Thr Leu 515520 525 Tyr Ser His Cys Thr Arg His Asp His Phe Ile Ser Ser Ser Leu Ser530 535 540 Ser Ser Ser Pro Ser Leu Phe Asp Gly Ala Val Ile Ser Thr ValThr 545 550 555 560 Thr Ala Thr Lys Lys His Phe Ser Ile Ile Leu Asn LeuLeu Gly Ile 565 570 575 Leu Leu Lys Lys Asp Asn Leu Asn Gln Asp Thr ArgLys Leu Leu Met 580 585 590 Thr Trp Ala Leu Glu Ala Ala Val Leu Met LysLys Ser Glu Thr Tyr 595 600 605 Ala Pro Leu Phe Ser Leu Pro Ser Phe HisLys Phe Cys Lys Gly Leu 610 615 620 Leu Ala Asn Thr Leu Val Glu Asp ValAsn Ile Cys Leu Gln Ala Cys 625 630 635 640 Ser Ser Leu His Ala Leu SerSer Ser Leu Pro Asp Asp Leu Leu Gln 645 650 655 Arg Cys Val Asp Val CysArg Val Gln Leu Val His Ser Gly Thr Arg 660 665 670 Ile Arg Gln Ala PheGly Lys Leu Leu Lys Ser Ile Pro Leu Asp Val 675 680 685 Val Leu Ser AsnAsn Asn His Thr Glu Ile Gln Glu Ile Ser Leu Ala 690 695 700 Leu Arg SerHis Met Ser Lys Ala Pro Ser Asn Thr Phe His Pro Gln 705 710 715 720 AspPhe Ser Asp Val Ile Ser Phe Ile Leu Tyr Gly Asn Ser His Arg 725 730 735Thr Gly Lys Asp Asn Trp Leu Glu Arg Leu Phe Tyr Ser Cys Gln Arg 740 745750 Leu Asp Lys Arg Asp Gln Ser Thr Ile Pro Arg Asn Leu Leu Lys Thr 755760 765 Asp Ala Val Leu Trp Gln Trp Ala Ile Trp Glu Ala Ala Gln Phe Thr770 775 780 Val Leu Ser Lys Leu Arg Thr Pro Leu Gly Arg Ala Gln Asp ThrPhe 785 790 795 800 Gln Thr Ile Glu Gly Ile Ile Arg Ser Leu Ala Ala HisThr Leu Asn 805 810 815 Pro Asp Gln Asp Val Ser Gln Trp Thr Thr Ala AspAsn Asp Glu Gly 820 825 830 His Gly Asn Asn Gln Leu Arg Leu Val Leu LeuLeu Gln Tyr Leu Glu 835 840 845 Asn Leu Glu Lys Leu Met Tyr Asn Ala TyrGlu Gly Cys Ala Asn Ala 850 855 860 Leu Thr Ser Pro Pro Lys Val Ile ArgThr Phe Phe Tyr Thr Asn Arg 865 870 875 880 Gln Thr Cys Gln Asp Trp LeuThr Arg Ile Arg Leu Ser Ile Met Arg 885 890 895 Val Gly Leu Leu Ala GlyGln Pro Ala Val Thr Val Arg His Gly Phe 900 905 910 Asp Leu Leu Thr GluMet Lys Thr Thr Ser Leu Ser Gln Gly Asn Glu 915 920 925 Leu Glu Val ThrIle Met Met Val Val Glu Ala Leu Cys Glu Leu His 930 935 940 Cys Pro GluAla Ile Gln Gly Ile Ala Val Trp Ser Ser Ser Ile Val 945 950 955 960 GlyLys Asn Leu Leu Trp Ile Asn Ser Val Ala Gln Gln Ala Glu Gly 965 970 975Arg Phe Glu Lys Ala Ser Val Glu Tyr Gln Glu His Leu Cys Ala Met 980 985990 Thr Gly Val Asp Cys Cys Ile Ser Ser Phe Asp Lys Ser Val Leu Thr 9951000 1005 Leu Ala Asn Ala Gly Arg Asn Ser Ala Ser Pro Lys His Ser LeuAsn 1010 1015 1020 Gly Glu Ser Arg Lys Thr Val Leu Ser Lys Pro Thr AspSer Ser Pro 1025 1030 1035 1040 Glu Val Ile Asn Tyr Leu Gly Asn Lys AlaCys Glu Cys Tyr Ile Ser 1045 1050 1055 Ile Ala Asp Trp Ala Ala Val GlnGlu Trp Gln Asn Ala Ile His Asp 1060 1065 1070 Leu Lys Lys Ser Thr SerSer Thr Ser Leu Asn Leu Lys Ala Asp Phe 1075 1080 1085 Asn Tyr Ile LysSer Leu Ser Ser Phe Glu Ser Gly Lys Phe Val Glu 1090 1095 1100 Cys ThrGlu Gln Leu Glu Leu Leu Pro Gly Glu Asn Ile Asn Leu Leu 1105 1110 11151120 Ala Gly Gly Ser Lys Glu Lys Ile Asp Met Lys Lys Leu Leu Pro Asn1125 1130 1135 Met Leu Ser Pro Asp Pro Arg Glu Leu Gln Lys Ser Ile GluVal Gln 1140 1145 1150 Leu Leu Arg Ser Ser Val Cys Leu Ala Thr Ala LeuAsn Pro Ile Glu 1155 1160 1165 Gln Asp Gln Lys Trp Gln Ser Ile Thr GluAsn Val Val Lys Tyr Leu 1170 1175 1180 Lys Gln Thr Ser Arg Ile Ala IleGly Pro Leu Arg Leu Ser Thr Leu 1185 1190 1195 1200 Thr Val Ser Gln SerLeu Pro Val Leu Ser Thr Leu Gln Leu Tyr Cys 1205 1210 1215 Ser Ser AlaLeu Glu Asn Thr Val Ser Asn Arg Leu Ser Thr Glu Asp 1220 1225 1230 CysLeu Ile Pro Leu Phe Ser Glu Ala Leu Arg Ser Cys Lys Gln His 1235 12401245 Asp Val Arg Pro Trp Met Gln Ala Leu Arg Tyr Thr Met Tyr Gln Asn1250 1255 1260 Gln Leu Leu Glu Lys Ile Lys Glu Gln Thr Val Pro Ile ArgSer His 1265 1270 1275 1280 Leu Met Glu Leu Gly Leu Thr Ala Ala Lys PheAla Arg Lys Arg Gly 1285 1290 1295 Asn Val Ser Leu Ala Thr Arg Leu LeuAla Gln Cys Ser Glu Val Gln 1300 1305 1310 Leu Gly Lys Thr Thr Thr AlaGln Asp Leu Val Gln His Phe Lys Lys 1315 1320 1325 Leu Ser Thr Gln GlyGln Val Asp Glu Lys Trp Gly Pro Glu Leu Asp 1330 1335 1340 Ile Glu LysThr Lys Leu Leu Tyr Thr Ala Gly Gln Ser Thr His Ala 1345 1350 1355 1360Met Glu Met Leu Ser Ser Cys Ala Ile Ser Phe Cys Lys Ser Val Lys 13651370 1375 Ala Glu Tyr Ala Val Ala Lys Ser Ile Leu Thr Leu Ala Lys TrpIle 1380 1385 1390 Gln Ala Glu Trp Lys Glu Ile Ser Gly Gln Leu Lys GlnVal Tyr Arg 1395 1400 1405 Ala Gln His Gln Gln Asn Phe Thr Gly Leu SerThr Leu Ser Lys Asn 1410 1415 1420 Ile Leu Thr Leu Ile Glu Leu Pro SerVal Asn Thr Met Glu Glu Glu 1425 1430 1435 1440 Tyr Pro Arg Ile Glu SerGlu Ser Thr Val His Ile Gly Val Gly Glu 1445 1450 1455 Pro Asp Phe IleLeu Gly Gln Leu Tyr His Leu Ser Ser Val Gln Ala 1460 1465 1470 Pro GluVal Ala Lys Ser Trp Ala Ala Leu Ala Ser Trp Ala Tyr Arg 1475 1480 1485Trp Gly Arg Lys Val Val Asp Asn Ala Ser Gln Gly Glu Gly Val Arg 14901495 1500 Leu Leu Pro Arg Glu Lys Ser Glu Val Gln Asn Leu Leu Pro AspThr 1505 1510 1515 1520 Ile Thr Glu Glu Glu Lys Glu Arg Ile Tyr Gly IleLeu Gly Gln Ala 1525 1530 1535 Val Cys Arg Pro Ala Gly Ile Gln Asp GluAsp Ile Thr Leu Gln Ile 1540 1545 1550 Thr Glu Ser Glu Asp Asn Glu GluAsp Asp Met Val Asp Val Ile Trp 1555 1560 1565 Arg Gln Leu Ile Ser SerCys Pro Trp Leu Ser Glu Leu Asp Glu Ser 1570 1575 1580 Ala Thr Glu GlyVal Ile Lys Val Trp Arg Lys Val Val Asp Arg Ile 1585 1590 1595 1600 PheSer Leu Tyr Lys Leu Ser Cys Ser Ala Tyr Phe Thr Phe Leu Lys 1605 16101615 Leu Asn Ala Gly Gln Ile Pro Leu Asp Glu Asp Asp Pro Arg Leu His1620 1625 1630 Leu Ser His Arg Val Glu Gln Ser Thr Asp Asp Met Ile ValMet Ala 1635 1640 1645 Thr Leu Arg Leu Leu Arg Leu Leu Val Lys His AlaGly Glu Leu Arg 1650 1655 1660 Gln Tyr Leu Glu His Gly Leu Glu Thr ThrPro Thr Ala Pro Trp Arg 1665 1670 1675 1680 Gly Ile Ile Pro Gln Leu PheSer Arg Leu Asn His Pro Glu Val Tyr 1685 1690 1695 Val Arg Gln Ser IleCys Asn Leu Leu Cys Arg Val Ala Gln Asp Ser 1700 1705 1710 Pro His LeuIle Leu Tyr Pro Ala Ile Val Gly Thr Ile Ser Leu Ser 1715 1720 1725 SerGlu Ser Gln Ala Ser Gly Asn Lys Phe Ser Thr Ala Ile Pro Thr 1730 17351740 Leu Leu Gly Asn Ile Gln Gly Glu Glu Leu Leu Val Ser Glu Cys Glu1745 1750 1755 1760 Gly Gly Ser Pro Pro Ala Ser Gln Asp Ser Asn Lys AspGlu Pro Lys 1765 1770 1775 Ser Gly Leu Asn Glu Asp Gln Ala Met Met GlnAsp Cys Tyr Ser Lys 1780 1785 1790 Ile Val Asp Lys Leu Ser Ser Ala AsnPro Thr Met Val Leu Gln Val 1795 1800 1805 Gln Met Leu Val Ala Glu LeuArg Arg Val Thr Val Leu Trp Asp Glu 1810 1815 1820 Leu Trp Leu Gly ValLeu Leu Gln Gln His Met Tyr Val Leu Arg Arg 1825 1830 1835 1840 Ile GlnGln Leu Glu Asp Glu Val Lys Arg Val Gln Asn Asn Asn Thr 1845 1850 1855Leu Arg Lys Glu Glu Lys Ile Ala Ile Met Arg Glu Lys His Thr Ala 18601865 1870 Leu Met Lys Pro Ile Val Phe Ala Leu Glu His Val Arg Ser IleThr 1875 1880 1885 Ala Ala Pro Ala Glu Thr Pro His Glu Lys Trp Phe GlnAsp Asn Tyr 1890 1895 1900 Gly Asp Ala Ile Glu Asn Ala Leu Glu Lys LeuLys Thr Pro Leu Asn 1905 1910 1915 1920 Pro Ala Lys Pro Gly Ser Ser TrpIle Pro Phe Lys Glu Ile Met Leu 1925 1930 1935 Ser Leu Gln Gln Arg AlaGln Lys Arg Ala Ser Tyr Ile Leu Arg Leu 1940 1945 1950 Glu Glu Ile SerPro Trp Leu Ala Ala Met Thr Asn Thr Glu Ile Ala 1955 1960 1965 Leu ProGly Glu Val Ser Ala Arg Asp Thr Val Thr Ile His Ser Val 1970 1975 1980Gly Gly Thr Ile Thr Ile Leu Pro Thr Lys Thr Lys Pro Lys Lys Leu 19851990 1995 2000 Leu Phe Leu Gly Ser Asp Gly Lys Ser Tyr Pro Tyr Leu PheLys Gly 2005 2010 2015 Leu Glu Asp Leu His Leu Asp Glu Arg Ile Met GlnPhe Leu Ser Ile 2020 2025 2030 Val Asn Thr Met Phe Ala Thr Ile Asn ArgGln Glu Thr Pro Arg Phe 2035 2040 2045 His Ala Arg His Tyr Ser Val ThrPro Leu Gly Thr Arg Ser Gly Leu 2050 2055 2060 Ile Gln Trp Val Asp GlyAla Thr Pro Leu Phe Gly Leu Tyr Lys Arg 2065 2070 2075 2080 Trp Gln GlnArg Glu Ala Ala Leu Gln Ala Gln Lys Ala Gln Asp Ser 2085 2090 2095 TyrGln Thr Pro Gln Asn Pro Gly Ile Val Pro Arg Pro Ser Glu Leu 2100 21052110 Tyr Tyr Ser Lys Ile Gly Pro Ala Leu Lys Thr Val Gly Leu Ser Leu2115 2120 2125 Asp Val Ser Arg Arg Asp Trp Pro Leu His Val Met Lys AlaVal Leu 2130 2135 2140 Glu Glu Leu Met Glu Ala Thr Pro Pro Asn Leu LeuAla Lys Glu Leu 2145 2150 2155 2160 Trp Ser Ser Cys Thr Thr Pro Asp GluTrp Trp Arg Val Thr Gln Ser 2165 2170 2175 Tyr Ala Arg Ser Thr Ala ValMet Ser Met Val Gly Tyr Ile Ile Gly 2180 2185 2190 Leu Gly Asp Arg HisLeu Asp Asn Val Leu Ile Asp Met Thr Thr Gly 2195 2200 2205 Glu Val ValHis Ile Asp Tyr Asn Val Cys Phe Glu Lys Gly Lys Ser 2210 2215 2220 LeuArg Val Pro Glu Lys Val Pro Phe Arg Met Thr Gln Asn Ile Glu 2225 22302235 2240 Thr Ala Leu Gly Val Thr Gly Val Glu Gly Val Phe Arg Leu SerCys 2245 2250 2255 Glu Gln Val Leu His Ile Met Arg Arg Gly Arg Glu ThrLeu Leu Thr 2260 2265 2270 Leu Leu Glu Ala Phe Val Tyr Asp Pro Leu ValAsp Trp Thr Ala Gly 2275 2280 2285 Gly Glu Ala Gly Phe Ala Gly Ala ValTyr Gly Gly Gly Gly Gln Gln 2290 2295 2300 Ala Glu Ser Lys Gln Ser LysArg Glu Met Glu Arg Glu Ile Thr Arg 2305 2310 2315 2320 Ser Leu Phe SerSer Arg Val Ala Glu Ile Lys Val Asn Trp Phe Lys 2325 2330 2335 Asn ArgAsp Glu Met Leu Val Val Leu Pro Lys Leu Asp Gly Ser Leu 2340 2345 2350Asp Glu Tyr Leu Ser Leu Gln Glu Gln Leu Thr Asp Val Glu Lys Leu 23552360 2365 Gln Gly Lys Leu Leu Glu Glu Ile Glu Phe Leu Glu Gly Ala GluGly 2370 2375 2380 Val Asp His Pro Ser His Thr Leu Gln His Arg Tyr SerGlu His Thr 2385 2390 2395 2400 Gln Leu Gln Thr Gln Gln Arg Ala Val GlnGlu Ala Ile Gln Val Lys 2405 2410 2415 Leu Asn Glu Phe Glu Gln Trp IleThr His Tyr Gln Ala Ala Phe Asn 2420 2425 2430 Asn Leu Glu Ala Thr GlnLeu Ala Ser Leu Leu Gln Glu Ile Ser Thr 2435 2440 2445 Gln Met Asp LeuGly Pro Pro Ser Tyr Val Pro Ala Thr Ala Phe Leu 2450 2455 2460 Gln AsnAla Gly Gln Ala His Leu Ile Ser Gln Cys Glu Gln Leu Glu 2465 2470 24752480 Gly Glu Val Gly Ala Leu Leu Gln Gln Arg Arg Ser Val Leu Arg Gly2485 2490 2495 Cys Leu Glu Gln Leu His His Tyr Ala Thr Val Ala Leu GlnTyr Pro 2500 2505 2510 Lys Ala Ile Phe Gln Lys His Arg Ile Glu Gln TrpLys Thr Trp Met 2515 2520 2525 Glu Glu Leu Ile Cys Asn Thr Thr Val GluArg Cys Gln Glu Leu Tyr 2530 2535 2540 Arg Lys Tyr Glu Met Gln Tyr AlaPro Gln Pro Pro Pro Thr Val Cys 2545 2550 2555 2560 Gln Phe Ile Thr AlaThr Glu Met Thr Leu Gln Arg Tyr Ala Ala Asp 2565 2570 2575 Ile Asn SerArg Leu Ile Arg Gln Val Glu Arg Leu Lys Gln Glu Ala 2580 2585 2590 ValThr Val Pro Val Cys Glu Asp Gln Leu Lys Glu Ile Glu Arg Cys 2595 26002605 Ile Lys Val Phe Leu His Glu Asn Gly Glu Glu Gly Ser Leu Ser Leu2610 2615 2620 Ala Ser Val Ile Ile Ser Ala Leu Cys Thr Leu Thr Arg ArgAsn Leu 2625 2630 2635 2640 Met Met Glu Gly Ala Ala Ser Ser Ala Gly GluGln Leu Val Asp Leu 2645 2650 2655 Thr Ser Arg Asp Gly Ala Trp Phe LeuGlu Glu Leu Cys Ser Met Ser 2660 2665 2670 Gly Asn Val Thr Cys Leu ValGln Leu Leu Lys Gln Cys His Leu Val 2675 2680 2685 Pro Gln Asp Leu AspIle Pro Asn Pro Met Glu Ala Ser Glu Thr Val 2690 2695 2700 His Leu AlaAsn Gly Val Tyr Thr Ser Leu Gln Glu Leu Asn Ser Asn 2705 2710 2715 2720Phe Arg Gln Ile Ile Phe Pro Glu Ala Leu Arg Cys Leu Met Lys Gly 27252730 2735 Glu Tyr Thr Leu Glu Ser Met Leu His Glu Leu Asp Gly Leu IleGlu 2740 2745 2750 Gln Thr Thr Asp Gly Val Pro Leu Gln Thr Leu Val GluSer Leu Gln 2755 2760 2765 Ala Tyr Leu Arg Asn Ala Ala Met Gly Leu GluGlu Glu Thr His Ala 2770 2775 2780 His Tyr Ile Asp Val Ala Arg Leu LeuHis Ala Gln Tyr Gly Glu Leu 2785 2790 2795 2800 Ile Gln Pro Arg Asn GlySer Val Asp Glu Thr Pro Lys Met Ser Ala 2805 2810 2815 Gly Gln Met LeuLeu Val Ala Phe Asp Gly Met Phe Ala Gln Val Glu 2820 2825 2830 Thr AlaPhe Ser Leu Leu Val Glu Lys Leu Asn Lys Met Glu Ile Pro 2835 2840 2845Ile Ala Trp Arg Lys Ile Asp Ile Ile Arg Glu Ala Arg Ser Thr Gln 28502855 2860 Val Asn Phe Phe Asp Asp Asp Asn His Arg Gln Val Leu Glu GluIle 2865 2870 2875 2880 Phe Phe Leu Lys Arg Leu Gln Thr Ile Lys Glu PhePhe Arg Leu Cys 2885 2890 2895 Gly Thr Phe Ser Lys Thr Leu Ser Gly SerSer Ser Leu Glu Asp Gln 2900 2905 2910 Asn Thr Val Asn Gly Pro Val GlnIle Val Asn Val Lys Thr Leu Phe 2915 2920 2925 Arg Asn Ser Cys Phe SerGlu Asp Gln Met Ala Lys Pro Ile Lys Ala 2930 2935 2940 Phe Thr Ala AspPhe Val Arg Gln Leu Leu Ile Gly Leu Pro Asn Gln 2945 2950 2955 2960 AlaLeu Gly Leu Thr Leu Cys Ser Phe Ile Ser Ala Leu Gly Val Asp 2965 29702975 Ile Ile Ala Gln Val Glu Ala Lys Asp Phe Gly Ala Glu Ser Lys Val2980 2985 2990 Ser Val Asp Asp Leu Cys Lys Lys Ala Val Glu His Asn IleGln Ile 2995 3000 3005 Gly Lys Phe Ser Gln Leu Val Met Asn Arg Ala ThrVal Leu Ala Ser 3010 3015 3020 Ser Tyr Asp Thr Ala Trp Lys Lys His AspLeu Val Arg Arg Leu Glu 3025 3030 3035 3040 Thr Ser Ile Ser Ser Cys LysThr Ser Leu Gln Arg Val Gln Leu His 3045 3050 3055 Ile Ala Met Phe GlnTrp Gln His Glu Asp Leu Leu Ile Asn Arg Pro 3060 3065 3070 Gln Ala MetSer Val Thr Pro Pro Pro Arg Ser Ala Ile Leu Thr Ser 3075 3080 3085 MetLys Lys Lys Leu His Thr Leu Ser Gln Ile Glu Thr Ser Ile Ala 3090 30953100 Thr Val Gln Glu Lys Leu Ala Ala Leu Glu Ser Ser Ile Glu Gln Arg3105 3110 3115 3120 Leu Lys Trp Ala Gly Gly Ala Asn Pro Ala Leu Ala ProVal Leu Gln 3125 3130 3135 Asp Phe Glu Ala Thr Ile Ala Glu Arg Arg AsnLeu Val Leu Lys Glu 3140 3145 3150 Ser Gln Arg Ala Ser Gln Val Thr PheLeu Cys Ser Asn Ile Ile His 3155 3160 3165 Phe Glu Ser Leu Arg Thr ArgThr Ala Glu Ala Leu Asn Leu Asp Ala 3170 3175 3180 Ala Leu Phe Glu LeuIle Lys Arg Cys Gln Gln Met Cys Ser Phe Ala 3185 3190 3195 3200 Ser GlnPhe Asn Ser Ser Val Ser Glu Leu Glu Leu Arg Leu Leu Gln 3205 3210 3215Arg Val Asp Thr Gly Leu Glu His Pro Ile Gly Ser Ser Glu Trp Leu 32203225 3230 Leu Ser Ala His Lys Gln Leu Thr Gln Asp Met Ser Thr Gln ArgAla 3235 3240 3245 Ile Gln Thr Glu Lys Glu Gln Gln Ile Glu Thr Val CysGlu Thr Ile 3250 3255 3260 Gln Asn Leu Val Asp Asn Ile Lys Thr Val LeuThr Gly His Asn Arg 3265 3270 3275 3280 Gln Leu Gly Asp Val Lys His LeuLeu Lys Ala Met Ala Lys Asp Glu 3285 3290 3295 Glu Ala Ala Leu Ala AspGly Glu Asp Val Pro Tyr Glu Asn Ser Val 3300 3305 3310 Arg Gln Phe LeuGly Glu Tyr Lys Ser Trp Gln Asp Asn Ile Gln Thr 3315 3320 3325 Val LeuPhe Thr Leu Val Gln Ala Met Gly Gln Val Arg Ser Gln Glu 3330 3335 3340His Val Glu Met Leu Gln Glu Ile Thr Pro Thr Leu Lys Glu Leu Lys 33453350 3355 3360 Thr Gln Ser Gln Ser Ile Tyr Asn Asn Leu Val Ser Phe AlaSer Pro 3365 3370 3375 Leu Val Thr Asp Ala Thr Asn Glu Cys Ser Ser ProThr Ser Ser Ala 3380 3385 3390 Thr Tyr Gln Pro Ser Phe Ala Ala Ala ValArg Ser Asn Thr Gly Gln 3395 3400 3405 Lys Thr Gln Pro Asp Val Met SerGln Asn Ala Arg Lys Leu Ile Gln 3410 3415 3420 Lys Asn Leu Ala Thr SerAla Asp Thr Pro Pro Ser Thr Val Pro Gly 3425 3430 3435 3440 Thr Gly LysSer Val Ala Cys Ser Pro Lys Lys Ala Val Arg Asp Pro 3445 3450 3455 LysThr Gly Lys Ala Val Gln Glu Arg Asn Ser Tyr Ala Val Ser Val 3460 34653470 Trp Lys Arg Val Lys Ala Lys Leu Glu Gly Arg Asp Val Asp Pro Asn3475 3480 3485 Arg Arg Met Ser Val Ala Glu Gln Val Asp Tyr Val Ile LysGlu Ala 3490 3495 3500 Thr Asn Leu Asp Asn Leu Ala Gln Leu Tyr Glu GlyTrp Thr Ala Trp 3505 3510 3515 3520 Val 3 2629 DNA Homo sapiens CDS(252)...(1433) 3 acttgtcatg gcgactgtcc agctttgtgc caggagcctc gcaggggttgatgggattgg 60 ggttttcccc tcccatgtgc tcaagactgg cgctaaaagt tttgagcttctcaaaagtct 120 agagccaccg tccagggagc aggtagctgc tgggctccgg ggacactttgcgttcgggct 180 gggagcgtgc tttccacgac ggtgacacgc ttccctggat tggcagccagactgccttcc 240 gggtcactgc c atg gag gag ccg cag tca gat cct agc gtc gagccc cct 290 Met Glu Glu Pro Gln Ser Asp Pro Ser Val Glu Pro Pro 1 5 10ctg agt cag gaa aca ttt tca gac cta tgg aaa cta ctt cct gaa aac 338 LeuSer Gln Glu Thr Phe Ser Asp Leu Trp Lys Leu Leu Pro Glu Asn 15 20 25 aacgtt ctg tcc ccc ttg ccg tcc caa gca atg gat gat ttg atg ctg 386 Asn ValLeu Ser Pro Leu Pro Ser Gln Ala Met Asp Asp Leu Met Leu 30 35 40 45 tccccg gac gat att gaa caa tgg ttc act gaa gac cca ggt cca gat 434 Ser ProAsp Asp Ile Glu Gln Trp Phe Thr Glu Asp Pro Gly Pro Asp 50 55 60 gaa gctccc aga atg cca gag gct gct ccc cgc gtg gcc cct gca cca 482 Glu Ala ProArg Met Pro Glu Ala Ala Pro Arg Val Ala Pro Ala Pro 65 70 75 gca gct cctaca ccg gcg gcc cct gca cca gcc ccc tcc tgg ccc ctg 530 Ala Ala Pro ThrPro Ala Ala Pro Ala Pro Ala Pro Ser Trp Pro Leu 80 85 90 tca tct tct gtccct tcc cag aaa acc tac cag ggc agc tac ggt ttc 578 Ser Ser Ser Val ProSer Gln Lys Thr Tyr Gln Gly Ser Tyr Gly Phe 95 100 105 cgt ctg ggc ttcttg cat tct ggg aca gcc aag tct gtg act tgc acg 626 Arg Leu Gly Phe LeuHis Ser Gly Thr Ala Lys Ser Val Thr Cys Thr 110 115 120 125 tac tcc cctgcc ctc aac aag atg ttt tgc caa ctg gcc aag acc tgc 674 Tyr Ser Pro AlaLeu Asn Lys Met Phe Cys Gln Leu Ala Lys Thr Cys 130 135 140 cct gtg cagctg tgg gtt gat tcc aca ccc ccg ccc ggc acc cgc gtc 722 Pro Val Gln LeuTrp Val Asp Ser Thr Pro Pro Pro Gly Thr Arg Val 145 150 155 cgc gcc atggcc atc tac aag cag tca cag cac atg acg gag gtt gtg 770 Arg Ala Met AlaIle Tyr Lys Gln Ser Gln His Met Thr Glu Val Val 160 165 170 agg cgc tgcccc cac cat gag cgc tgc tca gat agc gat ggt ctg gcc 818 Arg Arg Cys ProHis His Glu Arg Cys Ser Asp Ser Asp Gly Leu Ala 175 180 185 cct cct cagcat ctt atc cga gtg gaa gga aat ttg cgt gtg gag tat 866 Pro Pro Gln HisLeu Ile Arg Val Glu Gly Asn Leu Arg Val Glu Tyr 190 195 200 205 ttg gatgac aga aac act ttt cga cat agt gtg gtg gtg ccc tat gag 914 Leu Asp AspArg Asn Thr Phe Arg His Ser Val Val Val Pro Tyr Glu 210 215 220 ccg cctgag gtt ggc tct gac tgt acc acc atc cac tac aac tac atg 962 Pro Pro GluVal Gly Ser Asp Cys Thr Thr Ile His Tyr Asn Tyr Met 225 230 235 tgt aacagt tcc tgc atg ggc ggc atg aac cgg agg ccc atc ctc acc 1010 Cys Asn SerSer Cys Met Gly Gly Met Asn Arg Arg Pro Ile Leu Thr 240 245 250 atc atcaca ctg gaa gac tcc agt ggt aat cta ctg gga cgg aac agc 1058 Ile Ile ThrLeu Glu Asp Ser Ser Gly Asn Leu Leu Gly Arg Asn Ser 255 260 265 ttt gaggtg cgt gtt tgt gcc tgt cct ggg aga gac cgg cgc aca gag 1106 Phe Glu ValArg Val Cys Ala Cys Pro Gly Arg Asp Arg Arg Thr Glu 270 275 280 285 gaagag aat ctc cgc aag aaa ggg gag cct cac cac gag ctg ccc cca 1154 Glu GluAsn Leu Arg Lys Lys Gly Glu Pro His His Glu Leu Pro Pro 290 295 300 gggagc act aag cga gca ctg ccc aac aac acc agc tcc tct ccc cag 1202 Gly SerThr Lys Arg Ala Leu Pro Asn Asn Thr Ser Ser Ser Pro Gln 305 310 315 ccaaag aag aaa cca ctg gat gga gaa tat ttc acc ctt cag atc cgt 1250 Pro LysLys Lys Pro Leu Asp Gly Glu Tyr Phe Thr Leu Gln Ile Arg 320 325 330 gggcgt gag cgc ttc gag atg ttc cga gag ctg aat gag gcc ttg gaa 1298 Gly ArgGlu Arg Phe Glu Met Phe Arg Glu Leu Asn Glu Ala Leu Glu 335 340 345 ctcaag gat gcc cag gct ggg aag gag cca ggg ggg agc agg gct cac 1346 Leu LysAsp Ala Gln Ala Gly Lys Glu Pro Gly Gly Ser Arg Ala His 350 355 360 365tcc agc cac ctg aag tcc aaa aag ggt cag tct acc tcc cgc cat aaa 1394 SerSer His Leu Lys Ser Lys Lys Gly Gln Ser Thr Ser Arg His Lys 370 375 380aaa ctc atg ttc aag aca gaa ggg cct gac tca gac tga cattctccac 1443 LysLeu Met Phe Lys Thr Glu Gly Pro Asp Ser Asp * 385 390 ttcttgttccccactgacag cctcccaccc ccatctctcc ctcccctgcc attttgggtt 1503 ttgggtctttgaacccttgc ttgcaatagg tgtgcgtcag aagcacccag gacttccatt 1563 tgctttgtcccggggctcca ctgaacaagt tggcctgcac tggtgttttg ttgtggggag 1623 gaggatggggagtaggacat accagcttag attttaaggt ttttactgtg agggatgttt 1683 gggagatgtaagaaatgttc ttgcagttaa gggttagttt acaatcagcc acattctagg 1743 taggtaggggcccacttcac cgtactaacc agggaagctg tccctcatgt tgaattttct 1803 ctaacttcaaggcccatatc tgtgaaatgc tggcatttgc acctacctca cagagtgcat 1863 tgtgagggttaatgaaataa tgtacatctg gccttgaaac caccttttat tacatggggt 1923 ctaaaacttgacccccttga gggtgcctgt tccctctccc tctccctgtt ggctggtggg 1983 ttggtagtttctacagttgg gcagctggtt aggtagaggg agttgtcaag tcttgctggc 2043 ccagccaaaccctgtctgac aacctcttgg tcgaccttag tacctaaaag gaaatctcac 2103 cccatcccacaccctggagg atttcatctc ttgtatatga tgatctggat ccaccaagac 2163 ttgttttatgctcagggtca atttcttttt tctttttttt tttttttttt ctttttcttt 2223 gagactgggtctcgctttgt tgcccaggct ggagtggagt ggcgtgatct tggcttactg 2283 cagcctttgcctccccggct cgagcagtcc tgcctcagcc tccggagtag ctgggaccac 2343 aggttcatgccaccatggcc agccaacttt tgcatgtttt gtagagatgg ggtctcacag 2403 tgttgcccaggctggtctca aactcctggg ctcaggcgat ccacctgtct cagcctccca 2463 gagtgctgggattacaattg tgagccacca cgtggagctg gaagggtcaa catcttttac 2523 attctgcaagcacatctgca ttttcacccc acccttcccc tccttctccc tttttatatc 2583 ccatttttatatcgatctct tattttacaa taaaactttg ctgcca 2629 4 393 PRT Homo sapiens 4Met Glu Glu Pro Gln Ser Asp Pro Ser Val Glu Pro Pro Leu Ser Gln 1 5 1015 Glu Thr Phe Ser Asp Leu Trp Lys Leu Leu Pro Glu Asn Asn Val Leu 20 2530 Ser Pro Leu Pro Ser Gln Ala Met Asp Asp Leu Met Leu Ser Pro Asp 35 4045 Asp Ile Glu Gln Trp Phe Thr Glu Asp Pro Gly Pro Asp Glu Ala Pro 50 5560 Arg Met Pro Glu Ala Ala Pro Arg Val Ala Pro Ala Pro Ala Ala Pro 65 7075 80 Thr Pro Ala Ala Pro Ala Pro Ala Pro Ser Trp Pro Leu Ser Ser Ser 8590 95 Val Pro Ser Gln Lys Thr Tyr Gln Gly Ser Tyr Gly Phe Arg Leu Gly100 105 110 Phe Leu His Ser Gly Thr Ala Lys Ser Val Thr Cys Thr Tyr SerPro 115 120 125 Ala Leu Asn Lys Met Phe Cys Gln Leu Ala Lys Thr Cys ProVal Gln 130 135 140 Leu Trp Val Asp Ser Thr Pro Pro Pro Gly Thr Arg ValArg Ala Met 145 150 155 160 Ala Ile Tyr Lys Gln Ser Gln His Met Thr GluVal Val Arg Arg Cys 165 170 175 Pro His His Glu Arg Cys Ser Asp Ser AspGly Leu Ala Pro Pro Gln 180 185 190 His Leu Ile Arg Val Glu Gly Asn LeuArg Val Glu Tyr Leu Asp Asp 195 200 205 Arg Asn Thr Phe Arg His Ser ValVal Val Pro Tyr Glu Pro Pro Glu 210 215 220 Val Gly Ser Asp Cys Thr ThrIle His Tyr Asn Tyr Met Cys Asn Ser 225 230 235 240 Ser Cys Met Gly GlyMet Asn Arg Arg Pro Ile Leu Thr Ile Ile Thr 245 250 255 Leu Glu Asp SerSer Gly Asn Leu Leu Gly Arg Asn Ser Phe Glu Val 260 265 270 Arg Val CysAla Cys Pro Gly Arg Asp Arg Arg Thr Glu Glu Glu Asn 275 280 285 Leu ArgLys Lys Gly Glu Pro His His Glu Leu Pro Pro Gly Ser Thr 290 295 300 LysArg Ala Leu Pro Asn Asn Thr Ser Ser Ser Pro Gln Pro Lys Lys 305 310 315320 Lys Pro Leu Asp Gly Glu Tyr Phe Thr Leu Gln Ile Arg Gly Arg Glu 325330 335 Arg Phe Glu Met Phe Arg Glu Leu Asn Glu Ala Leu Glu Leu Lys Asp340 345 350 Ala Gln Ala Gly Lys Glu Pro Gly Gly Ser Arg Ala His Ser SerHis 355 360 365 Leu Lys Ser Lys Lys Gly Gln Ser Thr Ser Arg His Lys LysLeu Met 370 375 380 Phe Lys Thr Glu Gly Pro Asp Ser Asp 385 390 5 12606DNA Homo sapiens CDS (1703)...(10798) 5 attgacttga tgcacaacat cacaaaggcgatttttgaga actgatagtg catcagccga 60 cccaggtaat ttaaaatatt cttcatccagagatagaggt ggttcttcct cttacggact 120 gcaaccttca aattcagctg tggtgtctcggcaaaggcac gatgatacca gagtccacgc 180 tgacatacag aatgacgaaa aggagagatcgatgtcttat tgtgatgagt ctcgactgtc 240 gaatcttctt cggaggatca cccgggaagacgacagagac cgaagattgg ctactgtaaa 300 gcagttgaaa gaatttattc agcaaccagaaaataagctg gtactagtta aacaattgga 360 taatatcttg gctgctgtac atgacgtgcttaatgaaagt agcaaattgc ttcaggagtt 420 gagacaggag ggagcttgct gtcttggccttctttgtgct tctctgagct atgaggctga 480 gaagatcttc aagtggattt ttagcaaatttagctcatct gcaaaagatg aagttaaact 540 cctctactta tgtgccacct acaaagcactagagactgta ggagaaaaga aagccttttc 600 atctgtaatg cagcttgtaa tgaccagcctgcagtctatt cttgaaaatg tggatacacc 660 agaattgctt tgtaaatgtg ttaagtgcattcttttggtg gctcgatgtt accctcatat 720 tttcagcact aattttaggg atacagttgatatattagtt ggatggcata tagatcatac 780 tcagaaacct tcgctcacgc agcaggtatctgggtggttg cagagtttgg agccattttg 840 ggtagctgat cttgcatttt ctactactcttcttggtcag tttctggaag acatggaagc 900 atatgctgag gacctcagcc atgtggcctctggggaatca gtggatgaag atgtccctcc 960 tccatcagtg tcattaccaa agctggctgcacttctccgg gtatttagta ctgtggtgag 1020 gagcattggg gaacgcttca gcccaattcggggtcctcca attactgagg catatgtaac 1080 agatgttctg tacagagtaa tgagatgtgtgacggctgca aaccaggtgt ttttttctga 1140 ggctgtgttg acagctgcta atgagtgtgttggtgttttg ctcggcagct tggatcctag 1200 catgactata cattgtgaca tggtcattacatatggatta gaccaactgg agaattgcca 1260 gacttgtggt accgattata tcatctcagtcttgaattta ctcacgctga ttgttgaaca 1320 gataaatacg aaactgccat catcatttgtagaaaaactg tttataccat catctaaact 1380 actattcttg cgttatcata aagaaaaagaggttgttgct gtagcccatg ctgtttatca 1440 agcagtgctc agcttgaaga atattcctgttttggagact gcctataagt taatattggg 1500 agaaatgact tgtgccctaa acaacctcctacacagtcta caacttcctg aggcctgttc 1560 tgaaataaaa catgaggctt ttaagaatcatgtgttcaat gtagacaatg caaaatttgt 1620 agttatattt gacctcaatt gactacaaaatttgtagtta aatttgacta caattggaaa 1680 tgccaaaaac tcactaatag gg atg tgggcg cta tct cca act gtc ttt gca 1732 Met Trp Ala Leu Ser Pro Thr Val PheAla 1 5 10 ctt ctg agt aag aat ctg atg att gtg cac agt gac ctg gct gttcac 1780 Leu Leu Ser Lys Asn Leu Met Ile Val His Ser Asp Leu Ala Val His15 20 25 ttc cct gcc att cag tat gct gtg ctc tac aca ttg tat tct cat tgt1828 Phe Pro Ala Ile Gln Tyr Ala Val Leu Tyr Thr Leu Tyr Ser His Cys 3035 40 acc agg cat gat cac ttt atc tct agt agc ctc agt tct tcc tct cct1876 Thr Arg His Asp His Phe Ile Ser Ser Ser Leu Ser Ser Ser Ser Pro 4550 55 tct ttg ttt gat gga gct gtg att agc act gta act acg gct aca aag1924 Ser Leu Phe Asp Gly Ala Val Ile Ser Thr Val Thr Thr Ala Thr Lys 6065 70 aaa cat ttc tca att ata tta aat ctt ctg gga ata tta ctt aag aaa1972 Lys His Phe Ser Ile Ile Leu Asn Leu Leu Gly Ile Leu Leu Lys Lys 7580 85 90 gat aac ctt aac cag gac acg agg aaa ctg tta atg act tgg gct ttg2020 Asp Asn Leu Asn Gln Asp Thr Arg Lys Leu Leu Met Thr Trp Ala Leu 95100 105 gaa gca gct gtt tta atg aag aag tct gaa aca tac gca cct tta ttc2068 Glu Ala Ala Val Leu Met Lys Lys Ser Glu Thr Tyr Ala Pro Leu Phe 110115 120 tct ctt ccg tct ttc cat aaa ttt tgc aaa ggc ctt tta gcc aac act2116 Ser Leu Pro Ser Phe His Lys Phe Cys Lys Gly Leu Leu Ala Asn Thr 125130 135 ctc gtt gaa gat gtg aat atc tgt ctg cag gca tgc agc agt cta cat2164 Leu Val Glu Asp Val Asn Ile Cys Leu Gln Ala Cys Ser Ser Leu His 140145 150 gct ctg tcc tct tcc ttg cca gat gat ctt tta cag aga tgt gtc gat2212 Ala Leu Ser Ser Ser Leu Pro Asp Asp Leu Leu Gln Arg Cys Val Asp 155160 165 170 gtt tgc cgt gtt caa cta gtg cac agt gga act cgt att cga caagca 2260 Val Cys Arg Val Gln Leu Val His Ser Gly Thr Arg Ile Arg Gln Ala175 180 185 ttt gga aaa ctg ttg aaa tca att cct tta gat gtt gtc cta agcaat 2308 Phe Gly Lys Leu Leu Lys Ser Ile Pro Leu Asp Val Val Leu Ser Asn190 195 200 aac aat cac aca gaa att caa gaa att tct tta gca tta aga agtcac 2356 Asn Asn His Thr Glu Ile Gln Glu Ile Ser Leu Ala Leu Arg Ser His205 210 215 atg agt aaa gca cca agt aat aca ttc cac ccc caa gat ttc tctgat 2404 Met Ser Lys Ala Pro Ser Asn Thr Phe His Pro Gln Asp Phe Ser Asp220 225 230 gtt att agt ttt att ttg tat ggg aac tct cat aga aca ggg aaggac 2452 Val Ile Ser Phe Ile Leu Tyr Gly Asn Ser His Arg Thr Gly Lys Asp235 240 245 250 aat tgg ttg gaa aga ctg ttc tat agc tgc cag aga ctg gataag cgt 2500 Asn Trp Leu Glu Arg Leu Phe Tyr Ser Cys Gln Arg Leu Asp LysArg 255 260 265 gac cag tca aca att cca cgc aat ctc ctg aag aca gat gctgtc ctt 2548 Asp Gln Ser Thr Ile Pro Arg Asn Leu Leu Lys Thr Asp Ala ValLeu 270 275 280 tgg cag tgg gcc ata tgg gaa gct gca caa ttc act gtt ctttct aag 2596 Trp Gln Trp Ala Ile Trp Glu Ala Ala Gln Phe Thr Val Leu SerLys 285 290 295 ctg aga acc cca ctg ggc aga gct caa gac acc ttc cag acaatt gaa 2644 Leu Arg Thr Pro Leu Gly Arg Ala Gln Asp Thr Phe Gln Thr IleGlu 300 305 310 ggt atc att cga agt ctc gca gct cac aca tta aac cct gatcag gat 2692 Gly Ile Ile Arg Ser Leu Ala Ala His Thr Leu Asn Pro Asp GlnAsp 315 320 325 330 gtt agt cag tgg aca act gca gac aat gat gaa ggc catggt aac aac 2740 Val Ser Gln Trp Thr Thr Ala Asp Asn Asp Glu Gly His GlyAsn Asn 335 340 345 caa ctt aga ctt gtt ctt ctt ctg cag tat ctg gaa aatctg gag aaa 2788 Gln Leu Arg Leu Val Leu Leu Leu Gln Tyr Leu Glu Asn LeuGlu Lys 350 355 360 tta atg tat aat gca tac gag gga tgt gct aat gca ttaact tca cct 2836 Leu Met Tyr Asn Ala Tyr Glu Gly Cys Ala Asn Ala Leu ThrSer Pro 365 370 375 ccc aag gtc att aga act ttt ttc tat acc aat cgc caaact tgt cag 2884 Pro Lys Val Ile Arg Thr Phe Phe Tyr Thr Asn Arg Gln ThrCys Gln 380 385 390 gac tgg cta acg cgg att cga ctc tcc atc atg agg gtagga ttg ttg 2932 Asp Trp Leu Thr Arg Ile Arg Leu Ser Ile Met Arg Val GlyLeu Leu 395 400 405 410 gca ggc cag cct gca gtg aca gtg aga cat ggc tttgac ttg ctt aca 2980 Ala Gly Gln Pro Ala Val Thr Val Arg His Gly Phe AspLeu Leu Thr 415 420 425 gag atg aaa aca acc agc cta tct cag ggg aat gaattg gaa gta acc 3028 Glu Met Lys Thr Thr Ser Leu Ser Gln Gly Asn Glu LeuGlu Val Thr 430 435 440 att atg atg gtg gta gaa gca tta tgt gaa ctt cattgt cct gaa gct 3076 Ile Met Met Val Val Glu Ala Leu Cys Glu Leu His CysPro Glu Ala 445 450 455 ata cag gga att gct gtc tgg tca tca tct att gttgga aaa aat ctt 3124 Ile Gln Gly Ile Ala Val Trp Ser Ser Ser Ile Val GlyLys Asn Leu 460 465 470 ctg tgg att aac tca gtg gct caa cag gct gaa gggagg ttt gaa aag 3172 Leu Trp Ile Asn Ser Val Ala Gln Gln Ala Glu Gly ArgPhe Glu Lys 475 480 485 490 gcc tct gtg gag tac cag gaa cac ctg tgt gccatg aca ggt gtt gat 3220 Ala Ser Val Glu Tyr Gln Glu His Leu Cys Ala MetThr Gly Val Asp 495 500 505 tgc tgc atc tcc agc ttt gac aaa tcg gtg ctcacc tta gcc aat gct 3268 Cys Cys Ile Ser Ser Phe Asp Lys Ser Val Leu ThrLeu Ala Asn Ala 510 515 520 ggg cgt aac agt gcc agc ccg aaa cat tct ctgaat ggt gaa tcc aga 3316 Gly Arg Asn Ser Ala Ser Pro Lys His Ser Leu AsnGly Glu Ser Arg 525 530 535 aaa act gtg ctg tcc aaa ccg act gac tct tcccct gag gtt ata aat 3364 Lys Thr Val Leu Ser Lys Pro Thr Asp Ser Ser ProGlu Val Ile Asn 540 545 550 tat tta gga aat aaa gca tgt gag tgc tac atctca att gcc gat tgg 3412 Tyr Leu Gly Asn Lys Ala Cys Glu Cys Tyr Ile SerIle Ala Asp Trp 555 560 565 570 gct gct gtg cag gaa tgg cag aac gct atccat gac ttg aaa aag agt 3460 Ala Ala Val Gln Glu Trp Gln Asn Ala Ile HisAsp Leu Lys Lys Ser 575 580 585 acc agt agc act tcc ctc aac ctg aaa gctgac ttc aac tat ata aaa 3508 Thr Ser Ser Thr Ser Leu Asn Leu Lys Ala AspPhe Asn Tyr Ile Lys 590 595 600 tca tta agc agc ttt gag tct gga aaa tttgtt gaa tgt acc gag caa 3556 Ser Leu Ser Ser Phe Glu Ser Gly Lys Phe ValGlu Cys Thr Glu Gln 605 610 615 tta gaa ttg tta cca gga gaa aat atc aatcta ctt gct gga gga tca 3604 Leu Glu Leu Leu Pro Gly Glu Asn Ile Asn LeuLeu Ala Gly Gly Ser 620 625 630 aaa gaa aaa ata gac atg aaa aaa ctg cttcct aac atg tta agt ccg 3652 Lys Glu Lys Ile Asp Met Lys Lys Leu Leu ProAsn Met Leu Ser Pro 635 640 645 650 gat ccg agg gaa ctt cag aaa tcc attgaa gtt caa ttg tta aga agt 3700 Asp Pro Arg Glu Leu Gln Lys Ser Ile GluVal Gln Leu Leu Arg Ser 655 660 665 tct gtt tgt ttg gca act gct tta aacccg ata gaa caa gat cag aag 3748 Ser Val Cys Leu Ala Thr Ala Leu Asn ProIle Glu Gln Asp Gln Lys 670 675 680 tgg cag tct ata act gaa aat gtg gtaaag tac ttg aag caa aca tcc 3796 Trp Gln Ser Ile Thr Glu Asn Val Val LysTyr Leu Lys Gln Thr Ser 685 690 695 cgc atc gct att gga cct ctg aga ctttct act tta aca gtt tca cag 3844 Arg Ile Ala Ile Gly Pro Leu Arg Leu SerThr Leu Thr Val Ser Gln 700 705 710 tct ttg cca gtt cta agt acc ttg cagctg tat tgc tca tct gct ttg 3892 Ser Leu Pro Val Leu Ser Thr Leu Gln LeuTyr Cys Ser Ser Ala Leu 715 720 725 730 gag aac aca gtt tct aac aga ctttca aca gag gac tgt ctt att cca 3940 Glu Asn Thr Val Ser Asn Arg Leu SerThr Glu Asp Cys Leu Ile Pro 735 740 745 ctc ttc agt gaa gct tta cgt tcatgt aaa cag cat gac gtg agg cca 3988 Leu Phe Ser Glu Ala Leu Arg Ser CysLys Gln His Asp Val Arg Pro 750 755 760 tgg atg cag gca tta agg tat actatg tac cag aat cag ttg ttg gag 4036 Trp Met Gln Ala Leu Arg Tyr Thr MetTyr Gln Asn Gln Leu Leu Glu 765 770 775 aaa att aaa gaa caa aca gtc ccaatt aga agc cat ctc atg gaa tta 4084 Lys Ile Lys Glu Gln Thr Val Pro IleArg Ser His Leu Met Glu Leu 780 785 790 ggt cta aca gca gca aaa ttt gctaga aaa cga ggg aat gtg tcc ctt 4132 Gly Leu Thr Ala Ala Lys Phe Ala ArgLys Arg Gly Asn Val Ser Leu 795 800 805 810 gca aca aga ctg ctg gca cagtgc agt gaa gtt cag ctg gga aag acc 4180 Ala Thr Arg Leu Leu Ala Gln CysSer Glu Val Gln Leu Gly Lys Thr 815 820 825 acc act gca cag gat tta gtccaa cat ttt aaa aaa cta tca acc caa 4228 Thr Thr Ala Gln Asp Leu Val GlnHis Phe Lys Lys Leu Ser Thr Gln 830 835 840 ggt caa gtg gat gaa aaa tggggg ccc gaa ctt gat att gaa aaa acc 4276 Gly Gln Val Asp Glu Lys Trp GlyPro Glu Leu Asp Ile Glu Lys Thr 845 850 855 aaa ttg ctt tat aca gca ggccag tca aca cat gca atg gaa atg ttg 4324 Lys Leu Leu Tyr Thr Ala Gly GlnSer Thr His Ala Met Glu Met Leu 860 865 870 agt tct tgt gcc ata tct ttctgc aag tct gtg aaa gct gaa tat gca 4372 Ser Ser Cys Ala Ile Ser Phe CysLys Ser Val Lys Ala Glu Tyr Ala 875 880 885 890 gtt gct aaa tca att ctgaca ctg gct aaa tgg atc cag gca gaa tgg 4420 Val Ala Lys Ser Ile Leu ThrLeu Ala Lys Trp Ile Gln Ala Glu Trp 895 900 905 aaa gag att tca gga cagctg aaa cag gtt tac aga gct cag cac caa 4468 Lys Glu Ile Ser Gly Gln LeuLys Gln Val Tyr Arg Ala Gln His Gln 910 915 920 cag aac ttc aca ggt ctttct act ttg tct aaa aac ata ctc act cta 4516 Gln Asn Phe Thr Gly Leu SerThr Leu Ser Lys Asn Ile Leu Thr Leu 925 930 935 ata gaa ctg cca tct gttaat acg atg gaa gaa gag tat cct cgg atc 4564 Ile Glu Leu Pro Ser Val AsnThr Met Glu Glu Glu Tyr Pro Arg Ile 940 945 950 gag agt gaa tct aca gtgcat att gga gtt gga gaa cct gac ttc att 4612 Glu Ser Glu Ser Thr Val HisIle Gly Val Gly Glu Pro Asp Phe Ile 955 960 965 970 ttg gga cag ttg tatcac ctg tct tca gta cag gca cct gaa gta gcc 4660 Leu Gly Gln Leu Tyr HisLeu Ser Ser Val Gln Ala Pro Glu Val Ala 975 980 985 aaa tct tgg gca gcgttg gcc agc tgg gct tat agg tgg ggc aga aag 4708 Lys Ser Trp Ala Ala LeuAla Ser Trp Ala Tyr Arg Trp Gly Arg Lys 990 995 1000 gtg gtt gac aat gccagt cag gga gaa ggt gtt cgt ctg ctg cct aga 4756 Val Val Asp Asn Ala SerGln Gly Glu Gly Val Arg Leu Leu Pro Arg 1005 1010 1015 gaa aaa tct gaagtt cag aat cta ctt cca gac act ata act gag gaa 4804 Glu Lys Ser Glu ValGln Asn Leu Leu Pro Asp Thr Ile Thr Glu Glu 1020 1025 1030 gag aaa gagaga ata tat ggt att ctt gga cag gct gtg tgt cgg ccg 4852 Glu Lys Glu ArgIle Tyr Gly Ile Leu Gly Gln Ala Val Cys Arg Pro 1035 1040 1045 1050 gcgggg att cag gat gaa gat ata aca ctt cag ata act gag agt gaa 4900 Ala GlyIle Gln Asp Glu Asp Ile Thr Leu Gln Ile Thr Glu Ser Glu 1055 1060 1065gac aac gaa gaa gat gac atg gtt gat gtt atc tgg cgt cag ttg ata 4948 AspAsn Glu Glu Asp Asp Met Val Asp Val Ile Trp Arg Gln Leu Ile 1070 10751080 tca agc tgc cca tgg ctt tca gaa ctt gat gaa agt gca act gaa gga4996 Ser Ser Cys Pro Trp Leu Ser Glu Leu Asp Glu Ser Ala Thr Glu Gly1085 1090 1095 gtt att aaa gtg tgg agg aaa gtt gta gat aga ata ttc agcctg tac 5044 Val Ile Lys Val Trp Arg Lys Val Val Asp Arg Ile Phe Ser LeuTyr 1100 1105 1110 aaa ctc tct tgc agt gca tac ttt act ttc ctt aaa ctcaac gct ggt 5092 Lys Leu Ser Cys Ser Ala Tyr Phe Thr Phe Leu Lys Leu AsnAla Gly 1115 1120 1125 1130 caa att cct tta gat gag gat gac cct agg ctgcat tta agt cac aga 5140 Gln Ile Pro Leu Asp Glu Asp Asp Pro Arg Leu HisLeu Ser His Arg 1135 1140 1145 gtg gaa cag agc act gat gac atg att gtgatg gcc aca ttg cgc ctg 5188 Val Glu Gln Ser Thr Asp Asp Met Ile Val MetAla Thr Leu Arg Leu 1150 1155 1160 ctg cgg ttg ctc gtg aag cat gct ggtgag ctt cgg cag tat ctg gag 5236 Leu Arg Leu Leu Val Lys His Ala Gly GluLeu Arg Gln Tyr Leu Glu 1165 1170 1175 cac ggc ttg gag aca aca ccc actgca cca tgg aga gga att att ccg 5284 His Gly Leu Glu Thr Thr Pro Thr AlaPro Trp Arg Gly Ile Ile Pro 1180 1185 1190 caa ctt ttc tca cgc tta aaccac cct gaa gtg tat gtg cgc caa agt 5332 Gln Leu Phe Ser Arg Leu Asn HisPro Glu Val Tyr Val Arg Gln Ser 1195 1200 1205 1210 att tgt aac ctt ctctgc cgt gtg gct caa gat tcc cca cat ctc ata 5380 Ile Cys Asn Leu Leu CysArg Val Ala Gln Asp Ser Pro His Leu Ile 1215 1220 1225 ttg tat cct gcaata gtg ggt acc ata tcg ctt agt agt gaa tcc cag 5428 Leu Tyr Pro Ala IleVal Gly Thr Ile Ser Leu Ser Ser Glu Ser Gln 1230 1235 1240 gct tca ggaaat aaa ttt tcc act gca att cca act tta ctt ggc aat 5476 Ala Ser Gly AsnLys Phe Ser Thr Ala Ile Pro Thr Leu Leu Gly Asn 1245 1250 1255 att caagga gaa gaa ttg ctg gtt tct gaa tgt gag gga gga agt cct 5524 Ile Gln GlyGlu Glu Leu Leu Val Ser Glu Cys Glu Gly Gly Ser Pro 1260 1265 1270 cctgca tct cag gat agc aat aag gat gaa cct aaa agt gga tta aat 5572 Pro AlaSer Gln Asp Ser Asn Lys Asp Glu Pro Lys Ser Gly Leu Asn 1275 1280 12851290 gaa gac caa gcc atg atg cag gat tgt tac agc aaa att gta gat aag5620 Glu Asp Gln Ala Met Met Gln Asp Cys Tyr Ser Lys Ile Val Asp Lys1295 1300 1305 ctg tcc tct gca aac ccc acc atg gta tta cag gtt cag atgctc gtg 5668 Leu Ser Ser Ala Asn Pro Thr Met Val Leu Gln Val Gln Met LeuVal 1310 1315 1320 gct gaa ctg cgc agg gtc act gtg ctc tgg gat gag ctctgg ctg gga 5716 Ala Glu Leu Arg Arg Val Thr Val Leu Trp Asp Glu Leu TrpLeu Gly 1325 1330 1335 gtt ttg ctg caa caa cac atg tat gtc ctg aga cgaatt cag cag ctt 5764 Val Leu Leu Gln Gln His Met Tyr Val Leu Arg Arg IleGln Gln Leu 1340 1345 1350 gaa gat gag gtg aag aga gtc cag aac aac aacacc tta cgc aaa gaa 5812 Glu Asp Glu Val Lys Arg Val Gln Asn Asn Asn ThrLeu Arg Lys Glu 1355 1360 1365 1370 gag aaa att gca atc atg agg gag aagcac aca gct ttg atg aag ccc 5860 Glu Lys Ile Ala Ile Met Arg Glu Lys HisThr Ala Leu Met Lys Pro 1375 1380 1385 atc gta ttt gct ttg gag cat gtgagg agt atc aca gcg gct cct gca 5908 Ile Val Phe Ala Leu Glu His Val ArgSer Ile Thr Ala Ala Pro Ala 1390 1395 1400 gaa aca cct cat gaa aaa tggttt cag gat aac tat ggt gat gcc att 5956 Glu Thr Pro His Glu Lys Trp PheGln Asp Asn Tyr Gly Asp Ala Ile 1405 1410 1415 gaa aat gcc cta gaa aaactg aag act cca ttg aac cct gca aag cct 6004 Glu Asn Ala Leu Glu Lys LeuLys Thr Pro Leu Asn Pro Ala Lys Pro 1420 1425 1430 ggg agc agc tgg attcca ttt aaa gag ata atg cta agt ttg caa cag 6052 Gly Ser Ser Trp Ile ProPhe Lys Glu Ile Met Leu Ser Leu Gln Gln 1435 1440 1445 1450 aga gca cagaaa cgt gca agt tac atc ttg cgt ctt gaa gaa atc agt 6100 Arg Ala Gln LysArg Ala Ser Tyr Ile Leu Arg Leu Glu Glu Ile Ser 1455 1460 1465 cca tggttg gct gcc atg act aac act gaa att gct ctt cct ggg gaa 6148 Pro Trp LeuAla Ala Met Thr Asn Thr Glu Ile Ala Leu Pro Gly Glu 1470 1475 1480 gtctca gcc aga gac act gtc aca atc cat agt gtg ggc gga acc atc 6196 Val SerAla Arg Asp Thr Val Thr Ile His Ser Val Gly Gly Thr Ile 1485 1490 1495aca atc tta ccg act aaa acc aag cca aag aaa ctt ctc ttt ctt gga 6244 ThrIle Leu Pro Thr Lys Thr Lys Pro Lys Lys Leu Leu Phe Leu Gly 1500 15051510 tca gat ggg aag agc tat cct tat ctt ttc aaa gga ctg gag gat tta6292 Ser Asp Gly Lys Ser Tyr Pro Tyr Leu Phe Lys Gly Leu Glu Asp Leu1515 1520 1525 1530 cat ctg gat gag aga ata atg cag ttc cta tct att gtgaat acc atg 6340 His Leu Asp Glu Arg Ile Met Gln Phe Leu Ser Ile Val AsnThr Met 1535 1540 1545 ttt gct aca att aat cgc caa gaa aca ccc cgg ttccat gct cga cac 6388 Phe Ala Thr Ile Asn Arg Gln Glu Thr Pro Arg Phe HisAla Arg His 1550 1555 1560 tat tct gta aca cca cta gga aca aga tca ggacta atc cag tgg gta 6436 Tyr Ser Val Thr Pro Leu Gly Thr Arg Ser Gly LeuIle Gln Trp Val 1565 1570 1575 gat gga gcc aca ccc tta ttt ggt ctt tacaaa cga tgg caa caa cgg 6484 Asp Gly Ala Thr Pro Leu Phe Gly Leu Tyr LysArg Trp Gln Gln Arg 1580 1585 1590 gaa gct gcc tta caa gca caa aag gcccaa gat tcc tac caa act cct 6532 Glu Ala Ala Leu Gln Ala Gln Lys Ala GlnAsp Ser Tyr Gln Thr Pro 1595 1600 1605 1610 cag aat cct gga att gta ccccgt cct agt gaa ctt tat tac agt aaa 6580 Gln Asn Pro Gly Ile Val Pro ArgPro Ser Glu Leu Tyr Tyr Ser Lys 1615 1620 1625 att ggc cct gct ttg aaaaca gtt ggg ctt agc ctg gat gtg tcc cgt 6628 Ile Gly Pro Ala Leu Lys ThrVal Gly Leu Ser Leu Asp Val Ser Arg 1630 1635 1640 cgg gat tgg cct cttcat gta atg aag gca gta ttg gaa gag tta atg 6676 Arg Asp Trp Pro Leu HisVal Met Lys Ala Val Leu Glu Glu Leu Met 1645 1650 1655 gag gcc aca cccccg aat ctc ctt gcc aaa gag ctc tgg tca tct tgc 6724 Glu Ala Thr Pro ProAsn Leu Leu Ala Lys Glu Leu Trp Ser Ser Cys 1660 1665 1670 aca aca cctgat gaa tgg tgg aga gtt acg cag tct tat gca aga tct 6772 Thr Thr Pro AspGlu Trp Trp Arg Val Thr Gln Ser Tyr Ala Arg Ser 1675 1680 1685 1690 actgca gtc atg tct atg gtt gga tac ata att ggc ctt gga gac aga 6820 Thr AlaVal Met Ser Met Val Gly Tyr Ile Ile Gly Leu Gly Asp Arg 1695 1700 1705cat ctg gat aat gtt ctt ata gat atg acg act gga gaa gtt gtt cac 6868 HisLeu Asp Asn Val Leu Ile Asp Met Thr Thr Gly Glu Val Val His 1710 17151720 ata gat tac aat gtt tgc ttt gaa aaa ggt aaa agc ctt aga gtt cct6916 Ile Asp Tyr Asn Val Cys Phe Glu Lys Gly Lys Ser Leu Arg Val Pro1725 1730 1735 gag aaa gta cct ttt cga atg aca caa aac att gaa aca gcactg ggt 6964 Glu Lys Val Pro Phe Arg Met Thr Gln Asn Ile Glu Thr Ala LeuGly 1740 1745 1750 gta act gga gta gaa ggt gta ttt agg ctt tca tgt gagcag gtt tta 7012 Val Thr Gly Val Glu Gly Val Phe Arg Leu Ser Cys Glu GlnVal Leu 1755 1760 1765 1770 cac att atg cgg cgt ggc aga gag acc ctg ctgacg ctg ctg gag gcc 7060 His Ile Met Arg Arg Gly Arg Glu Thr Leu Leu ThrLeu Leu Glu Ala 1775 1780 1785 ttt gtg tac gac cct ctg gtg gac tgg acagca gga ggc gag gct ggg 7108 Phe Val Tyr Asp Pro Leu Val Asp Trp Thr AlaGly Gly Glu Ala Gly 1790 1795 1800 ttt gct ggt gct gtc tat ggt gga ggtggc cag cag gcc gag agc aag 7156 Phe Ala Gly Ala Val Tyr Gly Gly Gly GlyGln Gln Ala Glu Ser Lys 1805 1810 1815 cag agc aag aga gag atg gag cgagag atc acc cgc agc ctg ttt tct 7204 Gln Ser Lys Arg Glu Met Glu Arg GluIle Thr Arg Ser Leu Phe Ser 1820 1825 1830 tct aga gta gct gag att aaggtg aac tgg ttt aag aat aga gat gag 7252 Ser Arg Val Ala Glu Ile Lys ValAsn Trp Phe Lys Asn Arg Asp Glu 1835 1840 1845 1850 atg ctg gtt gtg cttccc aag ttg gac ggt agc tta gat gaa tac cta 7300 Met Leu Val Val Leu ProLys Leu Asp Gly Ser Leu Asp Glu Tyr Leu 1855 1860 1865 agc ttg caa gagcaa ctg aca gat gtg gaa aaa ctg cag ggc aaa cta 7348 Ser Leu Gln Glu GlnLeu Thr Asp Val Glu Lys Leu Gln Gly Lys Leu 1870 1875 1880 ctg gag gaaata gag ttt cta gaa gga gct gaa ggg gtg gat cat cct 7396 Leu Glu Glu IleGlu Phe Leu Glu Gly Ala Glu Gly Val Asp His Pro 1885 1890 1895 tct catact ctg caa cac agg tat tct gag cac acc caa cta cag act 7444 Ser His ThrLeu Gln His Arg Tyr Ser Glu His Thr Gln Leu Gln Thr 1900 1905 1910 cagcaa aga gct gtt cag gaa gca atc cag gtg aag ctg aat gaa ttt 7492 Gln GlnArg Ala Val Gln Glu Ala Ile Gln Val Lys Leu Asn Glu Phe 1915 1920 19251930 gaa caa tgg ata aca cat tat cag gct gca ttc aat aat tta gaa gca7540 Glu Gln Trp Ile Thr His Tyr Gln Ala Ala Phe Asn Asn Leu Glu Ala1935 1940 1945 aca cag ctt gca agc ttg ctt caa gag ata agc aca caa atggac ctt 7588 Thr Gln Leu Ala Ser Leu Leu Gln Glu Ile Ser Thr Gln Met AspLeu 1950 1955 1960 ggt cct cca agt tac gtg cca gca aca gcc ttt ctg cagaat gct ggt 7636 Gly Pro Pro Ser Tyr Val Pro Ala Thr Ala Phe Leu Gln AsnAla Gly 1965 1970 1975 cag gcc cac ttg att agc cag tgc gag cag ctg gagggg gag gtt ggt 7684 Gln Ala His Leu Ile Ser Gln Cys Glu Gln Leu Glu GlyGlu Val Gly 1980 1985 1990 gct ctc ctg cag cag agg cgc tcc gtg ctc cgtggc tgt ctg gag caa 7732 Ala Leu Leu Gln Gln Arg Arg Ser Val Leu Arg GlyCys Leu Glu Gln 1995 2000 2005 2010 ctg cat cac tat gca acc gtg gcc ctgcag tat ccg aag gcc ata ttt 7780 Leu His His Tyr Ala Thr Val Ala Leu GlnTyr Pro Lys Ala Ile Phe 2015 2020 2025 cag aaa cat cga att gaa cag tggaag acc tgg atg gaa gag ctc atc 7828 Gln Lys His Arg Ile Glu Gln Trp LysThr Trp Met Glu Glu Leu Ile 2030 2035 2040 tgt aac acc aca gta gag cgttgt caa gag ctc tat agg aaa tat gaa 7876 Cys Asn Thr Thr Val Glu Arg CysGln Glu Leu Tyr Arg Lys Tyr Glu 2045 2050 2055 atg caa tat gct ccc cagcca ccc cca aca gtg tgt cag ttc atc act 7924 Met Gln Tyr Ala Pro Gln ProPro Pro Thr Val Cys Gln Phe Ile Thr 2060 2065 2070 gcc act gaa atg accctg cag cga tac gca gca gac atc aac agc aga 7972 Ala Thr Glu Met Thr LeuGln Arg Tyr Ala Ala Asp Ile Asn Ser Arg 2075 2080 2085 2090 ctt att agacaa gtg gaa cgc ttg aaa cag gaa gct gtc act gtg cca 8020 Leu Ile Arg GlnVal Glu Arg Leu Lys Gln Glu Ala Val Thr Val Pro 2095 2100 2105 gtt tgtgaa gat cag ttg aaa gaa att gaa cgt tgc att aaa gtt ttc 8068 Val Cys GluAsp Gln Leu Lys Glu Ile Glu Arg Cys Ile Lys Val Phe 2110 2115 2120 cttcat gag aat gga gaa gaa gga tct ttg agt cta gca agt gtt att 8116 Leu HisGlu Asn Gly Glu Glu Gly Ser Leu Ser Leu Ala Ser Val Ile 2125 2130 2135att tct gcc ctt tgt acc ctt aca agg cgt aac ctg atg atg gaa ggt 8164 IleSer Ala Leu Cys Thr Leu Thr Arg Arg Asn Leu Met Met Glu Gly 2140 21452150 gca gcg tca agt gct gga gaa cag ctg gtt gat ctg act tct cgg gat8212 Ala Ala Ser Ser Ala Gly Glu Gln Leu Val Asp Leu Thr Ser Arg Asp2155 2160 2165 2170 gga gcc tgg ttc ttg gag gaa ctc tgc agt atg agc ggaaac gtc acc 8260 Gly Ala Trp Phe Leu Glu Glu Leu Cys Ser Met Ser Gly AsnVal Thr 2175 2180 2185 tgc ttg gtt cag tta ctg aag cag tgc cac ctg gtgcca cag gac tta 8308 Cys Leu Val Gln Leu Leu Lys Gln Cys His Leu Val ProGln Asp Leu 2190 2195 2200 gat atc ccg aac ccc atg gaa gcg tct gag acagtt cac tta gcc aat 8356 Asp Ile Pro Asn Pro Met Glu Ala Ser Glu Thr ValHis Leu Ala Asn 2205 2210 2215 gga gtg tat acc tca ctt cag gaa ttg aattcg aat ttc cgg caa atc 8404 Gly Val Tyr Thr Ser Leu Gln Glu Leu Asn SerAsn Phe Arg Gln Ile 2220 2225 2230 ata ttt cca gaa gca ctt cga tgt ttaatg aaa ggg gaa tac acg tta 8452 Ile Phe Pro Glu Ala Leu Arg Cys Leu MetLys Gly Glu Tyr Thr Leu 2235 2240 2245 2250 gaa agt atg ctg cat gaa ctggac ggt ctt att gag cag acc acc gat 8500 Glu Ser Met Leu His Glu Leu AspGly Leu Ile Glu Gln Thr Thr Asp 2255 2260 2265 ggc gtt ccc ctg cag actcta gtg gaa tct ctt cag gcc tac tta aga 8548 Gly Val Pro Leu Gln Thr LeuVal Glu Ser Leu Gln Ala Tyr Leu Arg 2270 2275 2280 aac gca gct atg ggactg gaa gaa gaa aca cat gct cat tac atc gat 8596 Asn Ala Ala Met Gly LeuGlu Glu Glu Thr His Ala His Tyr Ile Asp 2285 2290 2295 gtt gcc aga ctacta cat gct cag tac ggt gaa tta atc caa ccg aga 8644 Val Ala Arg Leu LeuHis Ala Gln Tyr Gly Glu Leu Ile Gln Pro Arg 2300 2305 2310 aat ggt tcagtt gat gaa aca ccc aaa atg tca gct ggc cag atg ctt 8692 Asn Gly Ser ValAsp Glu Thr Pro Lys Met Ser Ala Gly Gln Met Leu 2315 2320 2325 2330 ttggta gca ttc gat ggc atg ttt gct caa gtt gaa act gct ttc agc 8740 Leu ValAla Phe Asp Gly Met Phe Ala Gln Val Glu Thr Ala Phe Ser 2335 2340 2345tta tta gtt gaa aag ttg aac aag atg gaa att ccc ata gct tgg cga 8788 LeuLeu Val Glu Lys Leu Asn Lys Met Glu Ile Pro Ile Ala Trp Arg 2350 23552360 aag att gac atc ata agg gaa gcc agg agt act caa gtt aat ttt ttt8836 Lys Ile Asp Ile Ile Arg Glu Ala Arg Ser Thr Gln Val Asn Phe Phe2365 2370 2375 gat gat gat aat cac cgg cag gtg cta gaa gag att ttc tttcta aaa 8884 Asp Asp Asp Asn His Arg Gln Val Leu Glu Glu Ile Phe Phe LeuLys 2380 2385 2390 aga cta cag act att aag gag ttc ttc agg ctc tgt ggtacc ttt tct 8932 Arg Leu Gln Thr Ile Lys Glu Phe Phe Arg Leu Cys Gly ThrPhe Ser 2395 2400 2405 2410 aaa aca ttg tca gga tca agt tca ctt gaa gatcag aat act gtg aat 8980 Lys Thr Leu Ser Gly Ser Ser Ser Leu Glu Asp GlnAsn Thr Val Asn 2415 2420 2425 ggg cct gta cag att gtc aat gtg aaa accctt ttt aga aac tct tgt 9028 Gly Pro Val Gln Ile Val Asn Val Lys Thr LeuPhe Arg Asn Ser Cys 2430 2435 2440 ttc agt gaa gac caa atg gcc aaa cctatc aag gca ttc aca gct gac 9076 Phe Ser Glu Asp Gln Met Ala Lys Pro IleLys Ala Phe Thr Ala Asp 2445 2450 2455 ttt gtg agg cag ctc ttg ata gggcta ccc aac caa gcc ctc gga ctc 9124 Phe Val Arg Gln Leu Leu Ile Gly LeuPro Asn Gln Ala Leu Gly Leu 2460 2465 2470 aca ctg tgc agt ttt atc agtgct ctg ggt gta gac atc att gct caa 9172 Thr Leu Cys Ser Phe Ile Ser AlaLeu Gly Val Asp Ile Ile Ala Gln 2475 2480 2485 2490 gta gag gca aag gacttt ggt gcc gaa agc aaa gtt tct gtt gat gat 9220 Val Glu Ala Lys Asp PheGly Ala Glu Ser Lys Val Ser Val Asp Asp 2495 2500 2505 ctc tgt aag aaagcg gtg gaa cat aac atc cag ata ggg aag ttc tct 9268 Leu Cys Lys Lys AlaVal Glu His Asn Ile Gln Ile Gly Lys Phe Ser 2510 2515 2520 cag ctg gttatg aac agg gca act gtg tta gca agt tct tac gac act 9316 Gln Leu Val MetAsn Arg Ala Thr Val Leu Ala Ser Ser Tyr Asp Thr 2525 2530 2535 gcc tggaag aag cat gac ttg gtg cga agg cta gaa acc agt att tct 9364 Ala Trp LysLys His Asp Leu Val Arg Arg Leu Glu Thr Ser Ile Ser 2540 2545 2550 tcttgt aag aca agc ctg cag cgg gtt cag ctg cat att gcc atg ttt 9412 Ser CysLys Thr Ser Leu Gln Arg Val Gln Leu His Ile Ala Met Phe 2555 2560 25652570 cag tgg caa cat gaa gat cta ctt atc aat aga cca caa gcc atg tca9460 Gln Trp Gln His Glu Asp Leu Leu Ile Asn Arg Pro Gln Ala Met Ser2575 2580 2585 gtc aca cct ccc cca cgg tct gct atc cta acc agc atg aaaaag aag 9508 Val Thr Pro Pro Pro Arg Ser Ala Ile Leu Thr Ser Met Lys LysLys 2590 2595 2600 ctg cat acc ctg agc cag att gaa act tct att gca acagtt cag gag 9556 Leu His Thr Leu Ser Gln Ile Glu Thr Ser Ile Ala Thr ValGln Glu 2605 2610 2615 aag cta gct gca ctt gaa tca agt att gaa cag cgactc aag tgg gca 9604 Lys Leu Ala Ala Leu Glu Ser Ser Ile Glu Gln Arg LeuLys Trp Ala 2620 2625 2630 ggt ggt gcc aac cct gca ttg gcc cct gta ctacaa gat ttt gaa gca 9652 Gly Gly Ala Asn Pro Ala Leu Ala Pro Val Leu GlnAsp Phe Glu Ala 2635 2640 2645 2650 acg ata gct gaa aga aga aat ctt gtcctt aaa gag agc caa aga gca 9700 Thr Ile Ala Glu Arg Arg Asn Leu Val LeuLys Glu Ser Gln Arg Ala 2655 2660 2665 agt cag gtc aca ttt ctc tgc agcaat atc att cat ttt gaa agt tta 9748 Ser Gln Val Thr Phe Leu Cys Ser AsnIle Ile His Phe Glu Ser Leu 2670 2675 2680 cga aca aga act gca gaa gcctta aac ctg gat gcg gcg tta ttt gaa 9796 Arg Thr Arg Thr Ala Glu Ala LeuAsn Leu Asp Ala Ala Leu Phe Glu 2685 2690 2695 cta atc aag cga tgt cagcag atg tgt tcg ttt gca tca cag ttt aac 9844 Leu Ile Lys Arg Cys Gln GlnMet Cys Ser Phe Ala Ser Gln Phe Asn 2700 2705 2710 agt tca gtg tct gagtta gag ctt cgt tta tta cag aga gtg gac act 9892 Ser Ser Val Ser Glu LeuGlu Leu Arg Leu Leu Gln Arg Val Asp Thr 2715 2720 2725 2730 ggt ctt gaacat cct att ggc agc tct gaa tgg ctt ttg tca gca cac 9940 Gly Leu Glu HisPro Ile Gly Ser Ser Glu Trp Leu Leu Ser Ala His 2735 2740 2745 aaa cagttg acc cag gat atg tct act cag agg gca att cag aca gag 9988 Lys Gln LeuThr Gln Asp Met Ser Thr Gln Arg Ala Ile Gln Thr Glu 2750 2755 2760 aaagag cag cag ata gaa acg gtc tgt gaa aca att cag aat ctg gtt 10036 LysGlu Gln Gln Ile Glu Thr Val Cys Glu Thr Ile Gln Asn Leu Val 2765 27702775 gat aat ata aag act gtg ctc act ggt cat aac cga cag ctt gga gat10084 Asp Asn Ile Lys Thr Val Leu Thr Gly His Asn Arg Gln Leu Gly Asp2780 2785 2790 gtc aaa cat ctc ttg aaa gct atg gct aag gat gaa gaa gctgct ctg 10132 Val Lys His Leu Leu Lys Ala Met Ala Lys Asp Glu Glu AlaAla Leu 2795 2800 2805 2810 gca gat ggt gaa gat gtt ccc tat gag aac agtgtt agg cag ttt ttg 10180 Ala Asp Gly Glu Asp Val Pro Tyr Glu Asn SerVal Arg Gln Phe Leu 2815 2820 2825 ggt gaa tat aaa tca tgg caa gac aacatt caa aca gtt cta ttt aca 10228 Gly Glu Tyr Lys Ser Trp Gln Asp AsnIle Gln Thr Val Leu Phe Thr 2830 2835 2840 tta gtc cag gct atg ggt caggtt cga agt caa gaa cac gtt gaa atg 10276 Leu Val Gln Ala Met Gly GlnVal Arg Ser Gln Glu His Val Glu Met 2845 2850 2855 ctc cag gaa atc actccc acc ttg aaa gaa ctg aaa aca caa agt cag 10324 Leu Gln Glu Ile ThrPro Thr Leu Lys Glu Leu Lys Thr Gln Ser Gln 2860 2865 2870 agt atc tataat aat tta gtg agt ttt gca tca ccc tta gtc acc gat 10372 Ser Ile TyrAsn Asn Leu Val Ser Phe Ala Ser Pro Leu Val Thr Asp 2875 2880 2885 2890gca aca aat gaa tgt tcg agt cca acg tca tct gct act tat cag cca 10420Ala Thr Asn Glu Cys Ser Ser Pro Thr Ser Ser Ala Thr Tyr Gln Pro 28952900 2905 tcc ttc gct gca gca gtc cgg agt aac act ggc cag aag act cagcct 10468 Ser Phe Ala Ala Ala Val Arg Ser Asn Thr Gly Gln Lys Thr GlnPro 2910 2915 2920 gat gtc atg tca cag aat gct aga aag ctg atc cag aaaaat ctt gct 10516 Asp Val Met Ser Gln Asn Ala Arg Lys Leu Ile Gln LysAsn Leu Ala 2925 2930 2935 aca tca gct gat act cca cca agc acc gtt ccagga act ggc aag agt 10564 Thr Ser Ala Asp Thr Pro Pro Ser Thr Val ProGly Thr Gly Lys Ser 2940 2945 2950 gtt gct tgt agt cct aaa aag gca gtcaga gac cct aaa act ggg aaa 10612 Val Ala Cys Ser Pro Lys Lys Ala ValArg Asp Pro Lys Thr Gly Lys 2955 2960 2965 2970 gcg gtg caa gag aga aactcc tat gca gtg agt gtg tgg aag aga gtg 10660 Ala Val Gln Glu Arg AsnSer Tyr Ala Val Ser Val Trp Lys Arg Val 2975 2980 2985 aaa gcc aag ttagag ggc cga gat gtt gat ccg aat agg agg atg tca 10708 Lys Ala Lys LeuGlu Gly Arg Asp Val Asp Pro Asn Arg Arg Met Ser 2990 2995 3000 gtt gctgaa cag gtt gac tat gtc att aag gaa gca act aat cta gat 10756 Val AlaGlu Gln Val Asp Tyr Val Ile Lys Glu Ala Thr Asn Leu Asp 3005 3010 3015aac ttg gct cag ctg tat gaa ggt tgg aca gcc tgg gtg tga 10798 Asn LeuAla Gln Leu Tyr Glu Gly Trp Thr Ala Trp Val * 3020 3025 3030 atggcaagacagtagatgag tctggttaag cgaggtcaga catccaccag aatcaactca 10858 gcctcaggcatccaaagcca caccacagtc ggtggtgatg caactggggg cttactctga 10918 ggaaacctaggaaatctcgg tgcactagga agtgaatccc gcaggacagc tgcactcagg 10978 gatacgcccaacaccatggc ctgcaacccc agggtcaagg gtgaaggaaa gcaagctcac 11038 cgcctgaacacggagattgt ctttctgcca cagaacagca gcagacgtgt cgggaggtta 11098 gctgcggaaagaaatcggga tgccgcggag cacagagtga tttggaactc cattccacct 11158 gaccctgtgtgtacaatcca ggaaaaaaac aaaccccact cagaaacaga gaaaactggg 11218 gtcgcgaagaaatcacagcc aaggaagatt tgatgcattc agattctcgt gtaacacttg 11278 ttgcttggcaacagtactgg ttgggttgac cagtaagtag aaaaaggcta aaggctatgc 11338 gatatgaatttcagaaatgg actgaaaatg gagagctatg taacagatac actacagtag 11398 aagaacttacttctgaaatg aagggaaaaa aaccacccca tcgttcccta ctcctcccca 11458 ccacttacccgttccccctt tacctaatct agtagattag ccatctttca aattcacttt 11518 tatttcagtccttatatttc atatacttcc gtctcgatgc tgttaacaac ttctgataac 11578 atggaaaattcaaggattgt ttaaaggtct gatgatcaca cacaaaatgt aattccggtt 11638 atttaagtcatttctgtgat tctatcatgt acagtttcca gaattgtcac tgtgcattca 11698 aaagtaatgaatctaacaga catttgattt aatgtacact cccttttgct tatagtgtgc 11758 attttttttggaggtcattc aaattttccc tcttctgtga tagctgtagt ttctttcata 11818 gaaagtagctaatccagtgt aatcttttac ctttttaaaa accaagatag agtatctatt 11878 agagttttacattgttgatg atagattaac aataaagtga tgttctggtg gaggtagact 11938 gaaatttttttaattcatgt ttttcatttg atacttttaa tttacactta gtaaattaaa 11998 agttgtttaatttacttggc attttaggac atgtacatga aacagtgaaa atgagatcca 12058 ccaacatcttttattaagtt cagttattag tctgtgaagt gctttacttt ttgcacaatt 12118 ttaatagcttgctattcagt aatacattat agtgaattca tgatcaaggt ttccttaaat 12178 ttagcattgcatttcagtac tgactgtgta agctaaattg ctgatccaaa ataaaaaccc 12238 agactagaatagggttctta aaatcaagta tcaatacaaa atagaacaca attaaaatct 12298 taattgttggctgggcacag tggctcacgc ctgtaatccc agcactttgg gaggccgagg 12358 cgggcggatcatgaggttag gagagcgaga ccatcctggc taacacggtg aaaccccgtc 12418 tttactaaaatacaaaaaaa attagccggg tgtggtggcg ggcgcctgta gtcccagcta 12478 ctcgggaggctgaggcagga gaatggcgtg aacccaggag gcggagcttg cagtgagccg 12538 agattgtgccactgcactcc agcctgggca acagagctag actctgtgtc aaaaataaat 12598 gactagat12606 6 3031 PRT Homo sapiens 6 Met Trp Ala Leu Ser Pro Thr Val Phe AlaLeu Leu Ser Lys Asn Leu 1 5 10 15 Met Ile Val His Ser Asp Leu Ala ValHis Phe Pro Ala Ile Gln Tyr 20 25 30 Ala Val Leu Tyr Thr Leu Tyr Ser HisCys Thr Arg His Asp His Phe 35 40 45 Ile Ser Ser Ser Leu Ser Ser Ser SerPro Ser Leu Phe Asp Gly Ala 50 55 60 Val Ile Ser Thr Val Thr Thr Ala ThrLys Lys His Phe Ser Ile Ile 65 70 75 80 Leu Asn Leu Leu Gly Ile Leu LeuLys Lys Asp Asn Leu Asn Gln Asp 85 90 95 Thr Arg Lys Leu Leu Met Thr TrpAla Leu Glu Ala Ala Val Leu Met 100 105 110 Lys Lys Ser Glu Thr Tyr AlaPro Leu Phe Ser Leu Pro Ser Phe His 115 120 125 Lys Phe Cys Lys Gly LeuLeu Ala Asn Thr Leu Val Glu Asp Val Asn 130 135 140 Ile Cys Leu Gln AlaCys Ser Ser Leu His Ala Leu Ser Ser Ser Leu 145 150 155 160 Pro Asp AspLeu Leu Gln Arg Cys Val Asp Val Cys Arg Val Gln Leu 165 170 175 Val HisSer Gly Thr Arg Ile Arg Gln Ala Phe Gly Lys Leu Leu Lys 180 185 190 SerIle Pro Leu Asp Val Val Leu Ser Asn Asn Asn His Thr Glu Ile 195 200 205Gln Glu Ile Ser Leu Ala Leu Arg Ser His Met Ser Lys Ala Pro Ser 210 215220 Asn Thr Phe His Pro Gln Asp Phe Ser Asp Val Ile Ser Phe Ile Leu 225230 235 240 Tyr Gly Asn Ser His Arg Thr Gly Lys Asp Asn Trp Leu Glu ArgLeu 245 250 255 Phe Tyr Ser Cys Gln Arg Leu Asp Lys Arg Asp Gln Ser ThrIle Pro 260 265 270 Arg Asn Leu Leu Lys Thr Asp Ala Val Leu Trp Gln TrpAla Ile Trp 275 280 285 Glu Ala Ala Gln Phe Thr Val Leu Ser Lys Leu ArgThr Pro Leu Gly 290 295 300 Arg Ala Gln Asp Thr Phe Gln Thr Ile Glu GlyIle Ile Arg Ser Leu 305 310 315 320 Ala Ala His Thr Leu Asn Pro Asp GlnAsp Val Ser Gln Trp Thr Thr 325 330 335 Ala Asp Asn Asp Glu Gly His GlyAsn Asn Gln Leu Arg Leu Val Leu 340 345 350 Leu Leu Gln Tyr Leu Glu AsnLeu Glu Lys Leu Met Tyr Asn Ala Tyr 355 360 365 Glu Gly Cys Ala Asn AlaLeu Thr Ser Pro Pro Lys Val Ile Arg Thr 370 375 380 Phe Phe Tyr Thr AsnArg Gln Thr Cys Gln Asp Trp Leu Thr Arg Ile 385 390 395 400 Arg Leu SerIle Met Arg Val Gly Leu Leu Ala Gly Gln Pro Ala Val 405 410 415 Thr ValArg His Gly Phe Asp Leu Leu Thr Glu Met Lys Thr Thr Ser 420 425 430 LeuSer Gln Gly Asn Glu Leu Glu Val Thr Ile Met Met Val Val Glu 435 440 445Ala Leu Cys Glu Leu His Cys Pro Glu Ala Ile Gln Gly Ile Ala Val 450 455460 Trp Ser Ser Ser Ile Val Gly Lys Asn Leu Leu Trp Ile Asn Ser Val 465470 475 480 Ala Gln Gln Ala Glu Gly Arg Phe Glu Lys Ala Ser Val Glu TyrGln 485 490 495 Glu His Leu Cys Ala Met Thr Gly Val Asp Cys Cys Ile SerSer Phe 500 505 510 Asp Lys Ser Val Leu Thr Leu Ala Asn Ala Gly Arg AsnSer Ala Ser 515 520 525 Pro Lys His Ser Leu Asn Gly Glu Ser Arg Lys ThrVal Leu Ser Lys 530 535 540 Pro Thr Asp Ser Ser Pro Glu Val Ile Asn TyrLeu Gly Asn Lys Ala 545 550 555 560 Cys Glu Cys Tyr Ile Ser Ile Ala AspTrp Ala Ala Val Gln Glu Trp 565 570 575 Gln Asn Ala Ile His Asp Leu LysLys Ser Thr Ser Ser Thr Ser Leu 580 585 590 Asn Leu Lys Ala Asp Phe AsnTyr Ile Lys Ser Leu Ser Ser Phe Glu 595 600 605 Ser Gly Lys Phe Val GluCys Thr Glu Gln Leu Glu Leu Leu Pro Gly 610 615 620 Glu Asn Ile Asn LeuLeu Ala Gly Gly Ser Lys Glu Lys Ile Asp Met 625 630 635 640 Lys Lys LeuLeu Pro Asn Met Leu Ser Pro Asp Pro Arg Glu Leu Gln 645 650 655 Lys SerIle Glu Val Gln Leu Leu Arg Ser Ser Val Cys Leu Ala Thr 660 665 670 AlaLeu Asn Pro Ile Glu Gln Asp Gln Lys Trp Gln Ser Ile Thr Glu 675 680 685Asn Val Val Lys Tyr Leu Lys Gln Thr Ser Arg Ile Ala Ile Gly Pro 690 695700 Leu Arg Leu Ser Thr Leu Thr Val Ser Gln Ser Leu Pro Val Leu Ser 705710 715 720 Thr Leu Gln Leu Tyr Cys Ser Ser Ala Leu Glu Asn Thr Val SerAsn 725 730 735 Arg Leu Ser Thr Glu Asp Cys Leu Ile Pro Leu Phe Ser GluAla Leu 740 745 750 Arg Ser Cys Lys Gln His Asp Val Arg Pro Trp Met GlnAla Leu Arg 755 760 765 Tyr Thr Met Tyr Gln Asn Gln Leu Leu Glu Lys IleLys Glu Gln Thr 770 775 780 Val Pro Ile Arg Ser His Leu Met Glu Leu GlyLeu Thr Ala Ala Lys 785 790 795 800 Phe Ala Arg Lys Arg Gly Asn Val SerLeu Ala Thr Arg Leu Leu Ala 805 810 815 Gln Cys Ser Glu Val Gln Leu GlyLys Thr Thr Thr Ala Gln Asp Leu 820 825 830 Val Gln His Phe Lys Lys LeuSer Thr Gln Gly Gln Val Asp Glu Lys 835 840 845 Trp Gly Pro Glu Leu AspIle Glu Lys Thr Lys Leu Leu Tyr Thr Ala 850 855 860 Gly Gln Ser Thr HisAla Met Glu Met Leu Ser Ser Cys Ala Ile Ser 865 870 875 880 Phe Cys LysSer Val Lys Ala Glu Tyr Ala Val Ala Lys Ser Ile Leu 885 890 895 Thr LeuAla Lys Trp Ile Gln Ala Glu Trp Lys Glu Ile Ser Gly Gln 900 905 910 LeuLys Gln Val Tyr Arg Ala Gln His Gln Gln Asn Phe Thr Gly Leu 915 920 925Ser Thr Leu Ser Lys Asn Ile Leu Thr Leu Ile Glu Leu Pro Ser Val 930 935940 Asn Thr Met Glu Glu Glu Tyr Pro Arg Ile Glu Ser Glu Ser Thr Val 945950 955 960 His Ile Gly Val Gly Glu Pro Asp Phe Ile Leu Gly Gln Leu TyrHis 965 970 975 Leu Ser Ser Val Gln Ala Pro Glu Val Ala Lys Ser Trp AlaAla Leu 980 985 990 Ala Ser Trp Ala Tyr Arg Trp Gly Arg Lys Val Val AspAsn Ala Ser 995 1000 1005 Gln Gly Glu Gly Val Arg Leu Leu Pro Arg GluLys Ser Glu Val Gln 1010 1015 1020 Asn Leu Leu Pro Asp Thr Ile Thr GluGlu Glu Lys Glu Arg Ile Tyr 1025 1030 1035 1040 Gly Ile Leu Gly Gln AlaVal Cys Arg Pro Ala Gly Ile Gln Asp Glu 1045 1050 1055 Asp Ile Thr LeuGln Ile Thr Glu Ser Glu Asp Asn Glu Glu Asp Asp 1060 1065 1070 Met ValAsp Val Ile Trp Arg Gln Leu Ile Ser Ser Cys Pro Trp Leu 1075 1080 1085Ser Glu Leu Asp Glu Ser Ala Thr Glu Gly Val Ile Lys Val Trp Arg 10901095 1100 Lys Val Val Asp Arg Ile Phe Ser Leu Tyr Lys Leu Ser Cys SerAla 1105 1110 1115 1120 Tyr Phe Thr Phe Leu Lys Leu Asn Ala Gly Gln IlePro Leu Asp Glu 1125 1130 1135 Asp Asp Pro Arg Leu His Leu Ser His ArgVal Glu Gln Ser Thr Asp 1140 1145 1150 Asp Met Ile Val Met Ala Thr LeuArg Leu Leu Arg Leu Leu Val Lys 1155 1160 1165 His Ala Gly Glu Leu ArgGln Tyr Leu Glu His Gly Leu Glu Thr Thr 1170 1175 1180 Pro Thr Ala ProTrp Arg Gly Ile Ile Pro Gln Leu Phe Ser Arg Leu 1185 1190 1195 1200 AsnHis Pro Glu Val Tyr Val Arg Gln Ser Ile Cys Asn Leu Leu Cys 1205 12101215 Arg Val Ala Gln Asp Ser Pro His Leu Ile Leu Tyr Pro Ala Ile Val1220 1225 1230 Gly Thr Ile Ser Leu Ser Ser Glu Ser Gln Ala Ser Gly AsnLys Phe 1235 1240 1245 Ser Thr Ala Ile Pro Thr Leu Leu Gly Asn Ile GlnGly Glu Glu Leu 1250 1255 1260 Leu Val Ser Glu Cys Glu Gly Gly Ser ProPro Ala Ser Gln Asp Ser 1265 1270 1275 1280 Asn Lys Asp Glu Pro Lys SerGly Leu Asn Glu Asp Gln Ala Met Met 1285 1290 1295 Gln Asp Cys Tyr SerLys Ile Val Asp Lys Leu Ser Ser Ala Asn Pro 1300 1305 1310 Thr Met ValLeu Gln Val Gln Met Leu Val Ala Glu Leu Arg Arg Val 1315 1320 1325 ThrVal Leu Trp Asp Glu Leu Trp Leu Gly Val Leu Leu Gln Gln His 1330 13351340 Met Tyr Val Leu Arg Arg Ile Gln Gln Leu Glu Asp Glu Val Lys Arg1345 1350 1355 1360 Val Gln Asn Asn Asn Thr Leu Arg Lys Glu Glu Lys IleAla Ile Met 1365 1370 1375 Arg Glu Lys His Thr Ala Leu Met Lys Pro IleVal Phe Ala Leu Glu 1380 1385 1390 His Val Arg Ser Ile Thr Ala Ala ProAla Glu Thr Pro His Glu Lys 1395 1400 1405 Trp Phe Gln Asp Asn Tyr GlyAsp Ala Ile Glu Asn Ala Leu Glu Lys 1410 1415 1420 Leu Lys Thr Pro LeuAsn Pro Ala Lys Pro Gly Ser Ser Trp Ile Pro 1425 1430 1435 1440 Phe LysGlu Ile Met Leu Ser Leu Gln Gln Arg Ala Gln Lys Arg Ala 1445 1450 1455Ser Tyr Ile Leu Arg Leu Glu Glu Ile Ser Pro Trp Leu Ala Ala Met 14601465 1470 Thr Asn Thr Glu Ile Ala Leu Pro Gly Glu Val Ser Ala Arg AspThr 1475 1480 1485 Val Thr Ile His Ser Val Gly Gly Thr Ile Thr Ile LeuPro Thr Lys 1490 1495 1500 Thr Lys Pro Lys Lys Leu Leu Phe Leu Gly SerAsp Gly Lys Ser Tyr 1505 1510 1515 1520 Pro Tyr Leu Phe Lys Gly Leu GluAsp Leu His Leu Asp Glu Arg Ile 1525 1530 1535 Met Gln Phe Leu Ser IleVal Asn Thr Met Phe Ala Thr Ile Asn Arg 1540 1545 1550 Gln Glu Thr ProArg Phe His Ala Arg His Tyr Ser Val Thr Pro Leu 1555 1560 1565 Gly ThrArg Ser Gly Leu Ile Gln Trp Val Asp Gly Ala Thr Pro Leu 1570 1575 1580Phe Gly Leu Tyr Lys Arg Trp Gln Gln Arg Glu Ala Ala Leu Gln Ala 15851590 1595 1600 Gln Lys Ala Gln Asp Ser Tyr Gln Thr Pro Gln Asn Pro GlyIle Val 1605 1610 1615 Pro Arg Pro Ser Glu Leu Tyr Tyr Ser Lys Ile GlyPro Ala Leu Lys 1620 1625 1630 Thr Val Gly Leu Ser Leu Asp Val Ser ArgArg Asp Trp Pro Leu His 1635 1640 1645 Val Met Lys Ala Val Leu Glu GluLeu Met Glu Ala Thr Pro Pro Asn 1650 1655 1660 Leu Leu Ala Lys Glu LeuTrp Ser Ser Cys Thr Thr Pro Asp Glu Trp 1665 1670 1675 1680 Trp Arg ValThr Gln Ser Tyr Ala Arg Ser Thr Ala Val Met Ser Met 1685 1690 1695 ValGly Tyr Ile Ile Gly Leu Gly Asp Arg His Leu Asp Asn Val Leu 1700 17051710 Ile Asp Met Thr Thr Gly Glu Val Val His Ile Asp Tyr Asn Val Cys1715 1720 1725 Phe Glu Lys Gly Lys Ser Leu Arg Val Pro Glu Lys Val ProPhe Arg 1730 1735 1740 Met Thr Gln Asn Ile Glu Thr Ala Leu Gly Val ThrGly Val Glu Gly 1745 1750 1755 1760 Val Phe Arg Leu Ser Cys Glu Gln ValLeu His Ile Met Arg Arg Gly 1765 1770 1775 Arg Glu Thr Leu Leu Thr LeuLeu Glu Ala Phe Val Tyr Asp Pro Leu 1780 1785 1790 Val Asp Trp Thr AlaGly Gly Glu Ala Gly Phe Ala Gly Ala Val Tyr 1795 1800 1805 Gly Gly GlyGly Gln Gln Ala Glu Ser Lys Gln Ser Lys Arg Glu Met 1810 1815 1820 GluArg Glu Ile Thr Arg Ser Leu Phe Ser Ser Arg Val Ala Glu Ile 1825 18301835 1840 Lys Val Asn Trp Phe Lys Asn Arg Asp Glu Met Leu Val Val LeuPro 1845 1850 1855 Lys Leu Asp Gly Ser Leu Asp Glu Tyr Leu Ser Leu GlnGlu Gln Leu 1860 1865 1870 Thr Asp Val Glu Lys Leu Gln Gly Lys Leu LeuGlu Glu Ile Glu Phe 1875 1880 1885 Leu Glu Gly Ala Glu Gly Val Asp HisPro Ser His Thr Leu Gln His 1890 1895 1900 Arg Tyr Ser Glu His Thr GlnLeu Gln Thr Gln Gln Arg Ala Val Gln 1905 1910 1915 1920 Glu Ala Ile GlnVal Lys Leu Asn Glu Phe Glu Gln Trp Ile Thr His 1925 1930 1935 Tyr GlnAla Ala Phe Asn Asn Leu Glu Ala Thr Gln Leu Ala Ser Leu 1940 1945 1950Leu Gln Glu Ile Ser Thr Gln Met Asp Leu Gly Pro Pro Ser Tyr Val 19551960 1965 Pro Ala Thr Ala Phe Leu Gln Asn Ala Gly Gln Ala His Leu IleSer 1970 1975 1980 Gln Cys Glu Gln Leu Glu Gly Glu Val Gly Ala Leu LeuGln Gln Arg 1985 1990 1995 2000 Arg Ser Val Leu Arg Gly Cys Leu Glu GlnLeu His His Tyr Ala Thr 2005 2010 2015 Val Ala Leu Gln Tyr Pro Lys AlaIle Phe Gln Lys His Arg Ile Glu 2020 2025 2030 Gln Trp Lys Thr Trp MetGlu Glu Leu Ile Cys Asn Thr Thr Val Glu 2035 2040 2045 Arg Cys Gln GluLeu Tyr Arg Lys Tyr Glu Met Gln Tyr Ala Pro Gln 2050 2055 2060 Pro ProPro Thr Val Cys Gln Phe Ile Thr Ala Thr Glu Met Thr Leu 2065 2070 20752080 Gln Arg Tyr Ala Ala Asp Ile Asn Ser Arg Leu Ile Arg Gln Val Glu2085 2090 2095 Arg Leu Lys Gln Glu Ala Val Thr Val Pro Val Cys Glu AspGln Leu 2100 2105 2110 Lys Glu Ile Glu Arg Cys Ile Lys Val Phe Leu HisGlu Asn Gly Glu 2115 2120 2125 Glu Gly Ser Leu Ser Leu Ala Ser Val IleIle Ser Ala Leu Cys Thr 2130 2135 2140 Leu Thr Arg Arg Asn Leu Met MetGlu Gly Ala Ala Ser Ser Ala Gly 2145 2150 2155 2160 Glu Gln Leu Val AspLeu Thr Ser Arg Asp Gly Ala Trp Phe Leu Glu 2165 2170 2175 Glu Leu CysSer Met Ser Gly Asn Val Thr Cys Leu Val Gln Leu Leu 2180 2185 2190 LysGln Cys His Leu Val Pro Gln Asp Leu Asp Ile Pro Asn Pro Met 2195 22002205 Glu Ala Ser Glu Thr Val His Leu Ala Asn Gly Val Tyr Thr Ser Leu2210 2215 2220 Gln Glu Leu Asn Ser Asn Phe Arg Gln Ile Ile Phe Pro GluAla Leu 2225 2230 2235 2240 Arg Cys Leu Met Lys Gly Glu Tyr Thr Leu GluSer Met Leu His Glu 2245 2250 2255 Leu Asp Gly Leu Ile Glu Gln Thr ThrAsp Gly Val Pro Leu Gln Thr 2260 2265 2270 Leu Val Glu Ser Leu Gln AlaTyr Leu Arg Asn Ala Ala Met Gly Leu 2275 2280 2285 Glu Glu Glu Thr HisAla His Tyr Ile Asp Val Ala Arg Leu Leu His 2290 2295 2300 Ala Gln TyrGly Glu Leu Ile Gln Pro Arg Asn Gly Ser Val Asp Glu 2305 2310 2315 2320Thr Pro Lys Met Ser Ala Gly Gln Met Leu Leu Val Ala Phe Asp Gly 23252330 2335 Met Phe Ala Gln Val Glu Thr Ala Phe Ser Leu Leu Val Glu LysLeu 2340 2345 2350 Asn Lys Met Glu Ile Pro Ile Ala Trp Arg Lys Ile AspIle Ile Arg 2355 2360 2365 Glu Ala Arg Ser Thr Gln Val Asn Phe Phe AspAsp Asp Asn His Arg 2370 2375 2380 Gln Val Leu Glu Glu Ile Phe Phe LeuLys Arg Leu Gln Thr Ile Lys 2385 2390 2395 2400 Glu Phe Phe Arg Leu CysGly Thr Phe Ser Lys Thr Leu Ser Gly Ser 2405 2410 2415 Ser Ser Leu GluAsp Gln Asn Thr Val Asn Gly Pro Val Gln Ile Val 2420 2425 2430 Asn ValLys Thr Leu Phe Arg Asn Ser Cys Phe Ser Glu Asp Gln Met 2435 2440 2445Ala Lys Pro Ile Lys Ala Phe Thr Ala Asp Phe Val Arg Gln Leu Leu 24502455 2460 Ile Gly Leu Pro Asn Gln Ala Leu Gly Leu Thr Leu Cys Ser PheIle 2465 2470 2475 2480 Ser Ala Leu Gly Val Asp Ile Ile Ala Gln Val GluAla Lys Asp Phe 2485 2490 2495 Gly Ala Glu Ser Lys Val Ser Val Asp AspLeu Cys Lys Lys Ala Val 2500 2505 2510 Glu His Asn Ile Gln Ile Gly LysPhe Ser Gln Leu Val Met Asn Arg 2515 2520 2525 Ala Thr Val Leu Ala SerSer Tyr Asp Thr Ala Trp Lys Lys His Asp 2530 2535 2540 Leu Val Arg ArgLeu Glu Thr Ser Ile Ser Ser Cys Lys Thr Ser Leu 2545 2550 2555 2560 GlnArg Val Gln Leu His Ile Ala Met Phe Gln Trp Gln His Glu Asp 2565 25702575 Leu Leu Ile Asn Arg Pro Gln Ala Met Ser Val Thr Pro Pro Pro Arg2580 2585 2590 Ser Ala Ile Leu Thr Ser Met Lys Lys Lys Leu His Thr LeuSer Gln 2595 2600 2605 Ile Glu Thr Ser Ile Ala Thr Val Gln Glu Lys LeuAla Ala Leu Glu 2610 2615 2620 Ser Ser Ile Glu Gln Arg Leu Lys Trp AlaGly Gly Ala Asn Pro Ala 2625 2630 2635 2640 Leu Ala Pro Val Leu Gln AspPhe Glu Ala Thr Ile Ala Glu Arg Arg 2645 2650 2655 Asn Leu Val Leu LysGlu Ser Gln Arg Ala Ser Gln Val Thr Phe Leu 2660 2665 2670 Cys Ser AsnIle Ile His Phe Glu Ser Leu Arg Thr Arg Thr Ala Glu 2675 2680 2685 AlaLeu Asn Leu Asp Ala Ala Leu Phe Glu Leu Ile Lys Arg Cys Gln 2690 26952700 Gln Met Cys Ser Phe Ala Ser Gln Phe Asn Ser Ser Val Ser Glu Leu2705 2710 2715 2720 Glu Leu Arg Leu Leu Gln Arg Val Asp Thr Gly Leu GluHis Pro Ile 2725 2730 2735 Gly Ser Ser Glu Trp Leu Leu Ser Ala His LysGln Leu Thr Gln Asp 2740 2745 2750 Met Ser Thr Gln Arg Ala Ile Gln ThrGlu Lys Glu Gln Gln Ile Glu 2755 2760 2765 Thr Val Cys Glu Thr Ile GlnAsn Leu Val Asp Asn Ile Lys Thr Val 2770 2775 2780 Leu Thr Gly His AsnArg Gln Leu Gly Asp Val Lys His Leu Leu Lys 2785 2790 2795 2800 Ala MetAla Lys Asp Glu Glu Ala Ala Leu Ala Asp Gly Glu Asp Val 2805 2810 2815Pro Tyr Glu Asn Ser Val Arg Gln Phe Leu Gly Glu Tyr Lys Ser Trp 28202825 2830 Gln Asp Asn Ile Gln Thr Val Leu Phe Thr Leu Val Gln Ala MetGly 2835 2840 2845 Gln Val Arg Ser Gln Glu His Val Glu Met Leu Gln GluIle Thr Pro 2850 2855 2860 Thr Leu Lys Glu Leu Lys Thr Gln Ser Gln SerIle Tyr Asn Asn Leu 2865 2870 2875 2880 Val Ser Phe Ala Ser Pro Leu ValThr Asp Ala Thr Asn Glu Cys Ser 2885 2890 2895 Ser Pro Thr Ser Ser AlaThr Tyr Gln Pro Ser Phe Ala Ala Ala Val 2900 2905 2910 Arg Ser Asn ThrGly Gln Lys Thr Gln Pro Asp Val Met Ser Gln Asn 2915 2920 2925 Ala ArgLys Leu Ile Gln Lys Asn Leu Ala Thr Ser Ala Asp Thr Pro 2930 2935 2940Pro Ser Thr Val Pro Gly Thr Gly Lys Ser Val Ala Cys Ser Pro Lys 29452950 2955 2960 Lys Ala Val Arg Asp Pro Lys Thr Gly Lys Ala Val Gln GluArg Asn 2965 2970 2975 Ser Tyr Ala Val Ser Val Trp Lys Arg Val Lys AlaLys Leu Glu Gly 2980 2985 2990 Arg Asp Val Asp Pro Asn Arg Arg Met SerVal Ala Glu Gln Val Asp 2995 3000 3005 Tyr Val Ile Lys Glu Ala Thr AsnLeu Asp Asn Leu Ala Gln Leu Tyr 3010 3015 3020 Glu Gly Trp Thr Ala TrpVal 3025 3030 7 12539 DNA Homo sapiens CDS (141)...(10730) 7 gtggctacagtgtcaatgga ggatctgggg aaaatactta tggtcggaag tcgttggggc 60 aagagctgagggttaacaat gtgaccagcc ctgagttcac cagtgttcag catggcagtc 120 gtgctttagccaccaaagac atg agg aaa tca cag gag aga tcg atg tct tat 173 Met Arg LysSer Gln Glu Arg Ser Met Ser Tyr 1 5 10 tct gat gag tct cga ctg tcg aatctt ctt cgg agg atc acc cgg gaa 221 Ser Asp Glu Ser Arg Leu Ser Asn LeuLeu Arg Arg Ile Thr Arg Glu 15 20 25 gac gac aga gac cga aga ttg gct actgta aag cag ttg aaa gaa ttt 269 Asp Asp Arg Asp Arg Arg Leu Ala Thr ValLys Gln Leu Lys Glu Phe 30 35 40 att cag caa cca gaa aat aag ctg gta ctagtt aaa caa ttg gat aat 317 Ile Gln Gln Pro Glu Asn Lys Leu Val Leu ValLys Gln Leu Asp Asn 45 50 55 atc ttg gct gct gta cat gac gtg ctt aat gaaagt agc aaa ttg ctt 365 Ile Leu Ala Ala Val His Asp Val Leu Asn Glu SerSer Lys Leu Leu 60 65 70 75 cag gag ttg aga cag gag gga gct tgc tgt cttggc ctt ctt tgt gct 413 Gln Glu Leu Arg Gln Glu Gly Ala Cys Cys Leu GlyLeu Leu Cys Ala 80 85 90 tct ctg agc tat gag gct gag aag atc ttc aag tggatt ttt agc aaa 461 Ser Leu Ser Tyr Glu Ala Glu Lys Ile Phe Lys Trp IlePhe Ser Lys 95 100 105 ttt agc tca tct gca aaa gat gaa gtt aaa ctc ctctac tta tgt gcc 509 Phe Ser Ser Ser Ala Lys Asp Glu Val Lys Leu Leu TyrLeu Cys Ala 110 115 120 acc tac aaa gca cta gag act gta gga gaa aag aaagcc ttt tca tct 557 Thr Tyr Lys Ala Leu Glu Thr Val Gly Glu Lys Lys AlaPhe Ser Ser 125 130 135 gta atg cag ctt gta atg acc agc ctg cag tct attctt gaa aat gtg 605 Val Met Gln Leu Val Met Thr Ser Leu Gln Ser Ile LeuGlu Asn Val 140 145 150 155 gat aca cca gaa ttg ctt tgt aaa tgt gtt aagtgc att ctt ttg gtg 653 Asp Thr Pro Glu Leu Leu Cys Lys Cys Val Lys CysIle Leu Leu Val 160 165 170 gct cga tgt tac cct cat att ttc agc act aatttt agg gat aca gtt 701 Ala Arg Cys Tyr Pro His Ile Phe Ser Thr Asn PheArg Asp Thr Val 175 180 185 gat ata tta gtt gga tgg cat ata gat cat actcag aaa cct tcg ctc 749 Asp Ile Leu Val Gly Trp His Ile Asp His Thr GlnLys Pro Ser Leu 190 195 200 acg cag cag gta tct ggg tgg ttg cag agt ttggag cca ttt tgg gta 797 Thr Gln Gln Val Ser Gly Trp Leu Gln Ser Leu GluPro Phe Trp Val 205 210 215 gct gat ctt gca ttt tct act act ctt ctt ggtcag ttt ctg gaa gac 845 Ala Asp Leu Ala Phe Ser Thr Thr Leu Leu Gly GlnPhe Leu Glu Asp 220 225 230 235 atg gaa gca tat gct gag gac ctc agc catgtg gcc tct ggg gaa tca 893 Met Glu Ala Tyr Ala Glu Asp Leu Ser His ValAla Ser Gly Glu Ser 240 245 250 gtg gat gaa gat gtc cct cct cca tca gtgtca tta cca aag ctg gct 941 Val Asp Glu Asp Val Pro Pro Pro Ser Val SerLeu Pro Lys Leu Ala 255 260 265 gca ctt ctc cgg gta ttt agt act gtg gtgagg agc att ggg gaa cgc 989 Ala Leu Leu Arg Val Phe Ser Thr Val Val ArgSer Ile Gly Glu Arg 270 275 280 ttc agc cca att cgg ggt cct cca att actgag gca tat gta aca gat 1037 Phe Ser Pro Ile Arg Gly Pro Pro Ile Thr GluAla Tyr Val Thr Asp 285 290 295 gtt ctg tac aga gta atg aga tgt gtg acggct gca aac cag gtg ttt 1085 Val Leu Tyr Arg Val Met Arg Cys Val Thr AlaAla Asn Gln Val Phe 300 305 310 315 ttt tct gag gct gtg ttg aca gct gctaat gag tgt gtt ggt gtt ttg 1133 Phe Ser Glu Ala Val Leu Thr Ala Ala AsnGlu Cys Val Gly Val Leu 320 325 330 ctc ggc agc ttg gat cct agc atg actata cat tgt gac atg gtc att 1181 Leu Gly Ser Leu Asp Pro Ser Met Thr IleHis Cys Asp Met Val Ile 335 340 345 aca tat gga tta gac caa ctg gag aattgc cag act tgt ggt acc gat 1229 Thr Tyr Gly Leu Asp Gln Leu Glu Asn CysGln Thr Cys Gly Thr Asp 350 355 360 tat atc atc tca gtc ttg aat tta ctcacg ctg att gtt gaa cag ata 1277 Tyr Ile Ile Ser Val Leu Asn Leu Leu ThrLeu Ile Val Glu Gln Ile 365 370 375 aat acg aaa ctg cca tca tca ttt gtagaa aaa ctg ttt ata cca tca 1325 Asn Thr Lys Leu Pro Ser Ser Phe Val GluLys Leu Phe Ile Pro Ser 380 385 390 395 tct aaa cta cta ttc ttg cgt tatcat aaa gaa aaa gag gtt gtt gct 1373 Ser Lys Leu Leu Phe Leu Arg Tyr HisLys Glu Lys Glu Val Val Ala 400 405 410 gta gcc cat gct gtt tat caa gcagtg ctc agc ttg aag aat att cct 1421 Val Ala His Ala Val Tyr Gln Ala ValLeu Ser Leu Lys Asn Ile Pro 415 420 425 gtt ttg gag act gcc tat aag ttaata ttg gga gaa atg act tgt gcc 1469 Val Leu Glu Thr Ala Tyr Lys Leu IleLeu Gly Glu Met Thr Cys Ala 430 435 440 cta aac aac ctc cta cac agt ctacaa ctt cct gag gcc tgt tct gaa 1517 Leu Asn Asn Leu Leu His Ser Leu GlnLeu Pro Glu Ala Cys Ser Glu 445 450 455 ata aaa cat gag gct ttt aag aatcat gtg ttc aat gta gac aat gca 1565 Ile Lys His Glu Ala Phe Lys Asn HisVal Phe Asn Val Asp Asn Ala 460 465 470 475 aaa ttt gta gtt aaa ttt gacctc agt gcc ctg act aca att gga aat 1613 Lys Phe Val Val Lys Phe Asp LeuSer Ala Leu Thr Thr Ile Gly Asn 480 485 490 gcc aaa aac tca cta ata gggatg tgg gcg cta tct cca act gtc ttt 1661 Ala Lys Asn Ser Leu Ile Gly MetTrp Ala Leu Ser Pro Thr Val Phe 495 500 505 gca ctt ctg agt aag aat ctgatg att gtg cac agt gac ctg gct gtt 1709 Ala Leu Leu Ser Lys Asn Leu MetIle Val His Ser Asp Leu Ala Val 510 515 520 cac ttc cct gcc att cag tatgct gtg ctc tac aca ttg tat tct cat 1757 His Phe Pro Ala Ile Gln Tyr AlaVal Leu Tyr Thr Leu Tyr Ser His 525 530 535 tgt acc agg cat gat cac tttatc tct agt agc ctc agt tct gcc tct 1805 Cys Thr Arg His Asp His Phe IleSer Ser Ser Leu Ser Ser Ala Ser 540 545 550 555 cct tct ttg ttt gat ggagct gtg att agc act gta act acg gct aca 1853 Pro Ser Leu Phe Asp Gly AlaVal Ile Ser Thr Val Thr Thr Ala Thr 560 565 570 aag aaa cat ttc tca attata tta aat ctt ctg gga ata tta ctt aag 1901 Lys Lys His Phe Ser Ile IleLeu Asn Leu Leu Gly Ile Leu Leu Lys 575 580 585 aaa gat aac ctt aac caggac acg agg aaa ctg tta atg act tgg gct 1949 Lys Asp Asn Leu Asn Gln AspThr Arg Lys Leu Leu Met Thr Trp Ala 590 595 600 ttg gaa gca gct gtt ttaatg agg aag tct gaa aca tac gca cct tta 1997 Leu Glu Ala Ala Val Leu MetArg Lys Ser Glu Thr Tyr Ala Pro Leu 605 610 615 ttc tct ctt ccg tct ttccat aaa ttt tgc aaa ggc ctt tta gcc aac 2045 Phe Ser Leu Pro Ser Phe HisLys Phe Cys Lys Gly Leu Leu Ala Asn 620 625 630 635 act ctc gtt gaa gatgtg aat atc tgt ctg cag gca tgc agc agt cta 2093 Thr Leu Val Glu Asp ValAsn Ile Cys Leu Gln Ala Cys Ser Ser Leu 640 645 650 cat gct ctg tcc tcttcc ttg cca gat gat ctt tta cag aga tgt gtc 2141 His Ala Leu Ser Ser SerLeu Pro Asp Asp Leu Leu Gln Arg Cys Val 655 660 665 gat gtt tgc cgt gttcaa cta gtg cac agt gga act cgt att cga caa 2189 Asp Val Cys Arg Val GlnLeu Val His Ser Gly Thr Arg Ile Arg Gln 670 675 680 gca ttt gga aaa ctgttg aaa tca att cct tta gat gtt gtc cta agc 2237 Ala Phe Gly Lys Leu LeuLys Ser Ile Pro Leu Asp Val Val Leu Ser 685 690 695 aat aac aat cac acagaa att caa gaa att tct tta gca tta aga agt 2285 Asn Asn Asn His Thr GluIle Gln Glu Ile Ser Leu Ala Leu Arg Ser 700 705 710 715 cac atg agt aaagca cca agt aat aca ttc cac ccc caa gat ttc tct 2333 His Met Ser Lys AlaPro Ser Asn Thr Phe His Pro Gln Asp Phe Ser 720 725 730 gat gtt att agtttt att ttg tat ggg aac tct cat aga aca ggg aag 2381 Asp Val Ile Ser PheIle Leu Tyr Gly Asn Ser His Arg Thr Gly Lys 735 740 745 gac aat tgg ttggaa aga ctg ttc tat agc tgc cag aga ctg gat aag 2429 Asp Asn Trp Leu GluArg Leu Phe Tyr Ser Cys Gln Arg Leu Asp Lys 750 755 760 cgt gac cag tcaaca att cca cgc aat ctc ctg aag aca gat gct gtc 2477 Arg Asp Gln Ser ThrIle Pro Arg Asn Leu Leu Lys Thr Asp Ala Val 765 770 775 ctt tgg cag tgggcc ata tgg gaa gct gca caa ttc act gtt ctt tct 2525 Leu Trp Gln Trp AlaIle Trp Glu Ala Ala Gln Phe Thr Val Leu Ser 780 785 790 795 aag ctg agaacc cca ctg ggc aga gct caa gac acc ttc cag aca att 2573 Lys Leu Arg ThrPro Leu Gly Arg Ala Gln Asp Thr Phe Gln Thr Ile 800 805 810 gaa ggt atcatt cga agt ctc gca gct cac aca tta aac cct gat cag 2621 Glu Gly Ile IleArg Ser Leu Ala Ala His Thr Leu Asn Pro Asp Gln 815 820 825 gat gtt agtcag tgg aca act gca gac aat gat gaa ggc cat ggt aac 2669 Asp Val Ser GlnTrp Thr Thr Ala Asp Asn Asp Glu Gly His Gly Asn 830 835 840 aac caa cttaga ctt gtt ctt ctt ctg cag tat ctg gaa aat ctg gag 2717 Asn Gln Leu ArgLeu Val Leu Leu Leu Gln Tyr Leu Glu Asn Leu Glu 845 850 855 aaa tta atgtat aat gca tac gag gga tgt gct aat gca tta act tca 2765 Lys Leu Met TyrAsn Ala Tyr Glu Gly Cys Ala Asn Ala Leu Thr Ser 860 865 870 875 cct cccaag gtc att aga act ttt ttc tat acc aat cgc caa act tgt 2813 Pro Pro LysVal Ile Arg Thr Phe Phe Tyr Thr Asn Arg Gln Thr Cys 880 885 890 cag gactgg cta acg cgg att cga ctc tcc atc atg agg gta gga ttg 2861 Gln Asp TrpLeu Thr Arg Ile Arg Leu Ser Ile Met Arg Val Gly Leu 895 900 905 ttg gcaggc cag cct gca gtg aca gtg aga cat ggc ttt gac ttg ctt 2909 Leu Ala GlyGln Pro Ala Val Thr Val Arg His Gly Phe Asp Leu Leu 910 915 920 aca gagatg aaa aca acc agc cta tct cag ggg aat gaa ttg gaa gta 2957 Thr Glu MetLys Thr Thr Ser Leu Ser Gln Gly Asn Glu Leu Glu Val 925 930 935 acc attatg atg gtg gta gaa gca tta tgt gaa ctt cat tgt cct gaa 3005 Thr Ile MetMet Val Val Glu Ala Leu Cys Glu Leu His Cys Pro Glu 940 945 950 955 gctata cag gga att gct gtc tgg tca tca tct att gtt gga aaa aat 3053 Ala IleGln Gly Ile Ala Val Trp Ser Ser Ser Ile Val Gly Lys Asn 960 965 970 cttctg tgg att aac tca gtg gct caa cag gct gaa ggg agg ttt gaa 3101 Leu LeuTrp Ile Asn Ser Val Ala Gln Gln Ala Glu Gly Arg Phe Glu 975 980 985 aaggcc tct gtg gag tac cag gaa cac ctg tgt gcc atg aca ggt gtt 3149 Lys AlaSer Val Glu Tyr Gln Glu His Leu Cys Ala Met Thr Gly Val 990 995 1000 gattgc tgc atc tcc agc ttt gac aaa tcg gtg ctc acc tta gcc aat 3197 Asp CysCys Ile Ser Ser Phe Asp Lys Ser Val Leu Thr Leu Ala Asn 1005 1010 1015gct ggg cgt aac agt gcc agc ccg aaa cat tct ctg aat ggt gaa tcc 3245 AlaGly Arg Asn Ser Ala Ser Pro Lys His Ser Leu Asn Gly Glu Ser 1020 10251030 1035 aga aaa act gtg ctg tcc aaa ccg act gac tct tcc cct gag gttata 3293 Arg Lys Thr Val Leu Ser Lys Pro Thr Asp Ser Ser Pro Glu Val Ile1040 1045 1050 aat tat tta gga aat aaa gca tgt gag ttc tac atc tca attgcc gat 3341 Asn Tyr Leu Gly Asn Lys Ala Cys Glu Phe Tyr Ile Ser Ile AlaAsp 1055 1060 1065 tgg gct gct gtg cag gaa tgg cag aac gct atc cat gacttg aaa aag 3389 Trp Ala Ala Val Gln Glu Trp Gln Asn Ala Ile His Asp LeuLys Lys 1070 1075 1080 agt acc agt agc act tcc ctc aac ctg aaa gct gacttc aac tat ata 3437 Ser Thr Ser Ser Thr Ser Leu Asn Leu Lys Ala Asp PheAsn Tyr Ile 1085 1090 1095 aaa tca tta agc agc ttt gag tct gga aaa tttgtt gaa tgt acc gag 3485 Lys Ser Leu Ser Ser Phe Glu Ser Gly Lys Phe ValGlu Cys Thr Glu 1100 1105 1110 1115 cag tta gaa ttg tta cca gga gaa aatatc aat cta ctt gct gga gga 3533 Gln Leu Glu Leu Leu Pro Gly Glu Asn IleAsn Leu Leu Ala Gly Gly 1120 1125 1130 tca aaa gaa aaa ata gac atg aaaaaa ctg ctt cct aac atg tta agt 3581 Ser Lys Glu Lys Ile Asp Met Lys LysLeu Leu Pro Asn Met Leu Ser 1135 1140 1145 ccg gat ccg agg gaa ctt cagaaa tcc att gaa gtt caa ttg tta aga 3629 Pro Asp Pro Arg Glu Leu Gln LysSer Ile Glu Val Gln Leu Leu Arg 1150 1155 1160 agt tct gtt tgt ttg gcaact gct tta aac ccg ata gaa caa gat cag 3677 Ser Ser Val Cys Leu Ala ThrAla Leu Asn Pro Ile Glu Gln Asp Gln 1165 1170 1175 aag tgg cag tct ataact gaa aat gtg gta aag tac ttg aag caa aca 3725 Lys Trp Gln Ser Ile ThrGlu Asn Val Val Lys Tyr Leu Lys Gln Thr 1180 1185 1190 1195 tcc cgc atcgct att gga cct ctg aga ctt tct act tta aca gtt tca 3773 Ser Arg Ile AlaIle Gly Pro Leu Arg Leu Ser Thr Leu Thr Val Ser 1200 1205 1210 cag tctttg cca gtt cta agt acc ttg cag ctg tat tgc tca tct gct 3821 Gln Ser LeuPro Val Leu Ser Thr Leu Gln Leu Tyr Cys Ser Ser Ala 1215 1220 1225 ttggag aac aca gtt tct aac aga ctt tca aca gag gac tgt ctt att 3869 Leu GluAsn Thr Val Ser Asn Arg Leu Ser Thr Glu Asp Cys Leu Ile 1230 1235 1240cca ctc ttc agt gaa gct tta cgt tca tgt aaa cag cat gac gtg agg 3917 ProLeu Phe Ser Glu Ala Leu Arg Ser Cys Lys Gln His Asp Val Arg 1245 12501255 cca tgg atg cag gca tta agg tat act atg tac cag aat cag ttg ttg3965 Pro Trp Met Gln Ala Leu Arg Tyr Thr Met Tyr Gln Asn Gln Leu Leu1260 1265 1270 1275 gag aaa att aaa gaa caa aca gtc cca att aga agc catctc atg gaa 4013 Glu Lys Ile Lys Glu Gln Thr Val Pro Ile Arg Ser His LeuMet Glu 1280 1285 1290 tta ggt cta aca gca gca aaa ttt gct aga aaa cgaggg aat gtg tcc 4061 Leu Gly Leu Thr Ala Ala Lys Phe Ala Arg Lys Arg GlyAsn Val Ser 1295 1300 1305 ctt gca aca aga ctg ctg gca cag tgc agt gaagtt cag ctg gga aag 4109 Leu Ala Thr Arg Leu Leu Ala Gln Cys Ser Glu ValGln Leu Gly Lys 1310 1315 1320 acc acc act gca cag gat tta gtc caa catttt aaa aaa cta tca acc 4157 Thr Thr Thr Ala Gln Asp Leu Val Gln His PheLys Lys Leu Ser Thr 1325 1330 1335 caa ggt caa gtg gat gaa aaa tgg gggccc gaa ctt gat att gaa aaa 4205 Gln Gly Gln Val Asp Glu Lys Trp Gly ProGlu Leu Asp Ile Glu Lys 1340 1345 1350 1355 acc aaa ttg ctt tat aca gcaggc cag tca aca cat gca atg gaa atg 4253 Thr Lys Leu Leu Tyr Thr Ala GlyGln Ser Thr His Ala Met Glu Met 1360 1365 1370 ttg agt tct tgt gcc atatct ttc tgc aag tct gtg aaa gct gaa tat 4301 Leu Ser Ser Cys Ala Ile SerPhe Cys Lys Ser Val Lys Ala Glu Tyr 1375 1380 1385 gca gtt gct aaa tcaatt ctg aca ctg gct aaa tgg atc cag gca gaa 4349 Ala Val Ala Lys Ser IleLeu Thr Leu Ala Lys Trp Ile Gln Ala Glu 1390 1395 1400 tgg aaa gag atttca gga cag ctg aaa cag gtt tac aga gct cag cac 4397 Trp Lys Glu Ile SerGly Gln Leu Lys Gln Val Tyr Arg Ala Gln His 1405 1410 1415 caa cag aacttc aca ggt ctt tct act ttg tct aaa aac ata ctc act 4445 Gln Gln Asn PheThr Gly Leu Ser Thr Leu Ser Lys Asn Ile Leu Thr 1420 1425 1430 1435 ctaata gaa ctg cca tct gtt aat acg atg gaa gaa gag tat cct cgg 4493 Leu IleGlu Leu Pro Ser Val Asn Thr Met Glu Glu Glu Tyr Pro Arg 1440 1445 1450atc gag agt gaa tct aca gtg cat att gga gtt gga gaa cct gac ttc 4541 IleGlu Ser Glu Ser Thr Val His Ile Gly Val Gly Glu Pro Asp Phe 1455 14601465 att ttg gga cag ttg tat cac ctg tct tca gta cag gca cct gaa gta4589 Ile Leu Gly Gln Leu Tyr His Leu Ser Ser Val Gln Ala Pro Glu Val1470 1475 1480 gcc aaa tct tgg gca gcg ttg gcc agc tgg gct tat agg tggggc aga 4637 Ala Lys Ser Trp Ala Ala Leu Ala Ser Trp Ala Tyr Arg Trp GlyArg 1485 1490 1495 aag gtg gtt gac aat gcc agt cag gga gaa ggt gtt cgtctg ctg cct 4685 Lys Val Val Asp Asn Ala Ser Gln Gly Glu Gly Val Arg LeuLeu Pro 1500 1505 1510 1515 aga gaa aaa tct gaa gtt cag aat cta ctt ccagac act ata act gag 4733 Arg Glu Lys Ser Glu Val Gln Asn Leu Leu Pro AspThr Ile Thr Glu 1520 1525 1530 gaa gag aaa gag aga ata tat ggt att cttgga cag gct gtg tgt cgg 4781 Glu Glu Lys Glu Arg Ile Tyr Gly Ile Leu GlyGln Ala Val Cys Arg 1535 1540 1545 ccg gcg ggg att cag gat gaa gat ataaca ctt cag ata act gag agt 4829 Pro Ala Gly Ile Gln Asp Glu Asp Ile ThrLeu Gln Ile Thr Glu Ser 1550 1555 1560 gaa gac aac gaa gaa gat gac atggtt gat gtt atc tgg cgt cag ttg 4877 Glu Asp Asn Glu Glu Asp Asp Met ValAsp Val Ile Trp Arg Gln Leu 1565 1570 1575 ata tca agc tgc cca tgg ctttca gaa ctt gat gaa agt gca act gaa 4925 Ile Ser Ser Cys Pro Trp Leu SerGlu Leu Asp Glu Ser Ala Thr Glu 1580 1585 1590 1595 gga gtt att aaa gtgtgg agg aaa gtt gta gat aga ata ttc agc ctg 4973 Gly Val Ile Lys Val TrpArg Lys Val Val Asp Arg Ile Phe Ser Leu 1600 1605 1610 tac aaa ctc tcttgc agt gca tac ttt act ttc ctt aaa ctc aac gct 5021 Tyr Lys Leu Ser CysSer Ala Tyr Phe Thr Phe Leu Lys Leu Asn Ala 1615 1620 1625 ggt caa attcct tta gat gag gat gac cct agg ctg cat tta agt cac 5069 Gly Gln Ile ProLeu Asp Glu Asp Asp Pro Arg Leu His Leu Ser His 1630 1635 1640 aga gtggaa cag agc act gat gac atg att gtg atg gcc aca ttg cgc 5117 Arg Val GluGln Ser Thr Asp Asp Met Ile Val Met Ala Thr Leu Arg 1645 1650 1655 ctgctg cgg ttg ctc gtg aag cat gct ggt gag ctt cgg cag tat ctg 5165 Leu LeuArg Leu Leu Val Lys His Ala Gly Glu Leu Arg Gln Tyr Leu 1660 1665 16701675 gag cac ggc ttg gag aca aca ccc act gca cca tgg agg gga att att5213 Glu His Gly Leu Glu Thr Thr Pro Thr Ala Pro Trp Arg Gly Ile Ile1680 1685 1690 ccg caa ctt ttc tca cgc tta aac cac cct gaa gtg tat gtgcgc caa 5261 Pro Gln Leu Phe Ser Arg Leu Asn His Pro Glu Val Tyr Val ArgGln 1695 1700 1705 agt att tgt aac ctt ctc tgc cgt gtg gct caa gat tcccca cat ctc 5309 Ser Ile Cys Asn Leu Leu Cys Arg Val Ala Gln Asp Ser ProHis Leu 1710 1715 1720 ata ttg tat cct gca ata gtg ggt acc ata tcg cttagt agt gaa tcc 5357 Ile Leu Tyr Pro Ala Ile Val Gly Thr Ile Ser Leu SerSer Glu Ser 1725 1730 1735 cag gct tca gga aat aaa ttt tcc act gca attcca act tta ctt ggc 5405 Gln Ala Ser Gly Asn Lys Phe Ser Thr Ala Ile ProThr Leu Leu Gly 1740 1745 1750 1755 aat att caa gga gaa gaa ttg ctg gtttct gaa tgt gag gga gga agt 5453 Asn Ile Gln Gly Glu Glu Leu Leu Val SerGlu Cys Glu Gly Gly Ser 1760 1765 1770 cct cct gca tct cag gat agc aataag gat gaa cct aaa agt gga tta 5501 Pro Pro Ala Ser Gln Asp Ser Asn LysAsp Glu Pro Lys Ser Gly Leu 1775 1780 1785 aat gaa gac caa gcc atg atgcag gat tgt tac agc aaa att gta gat 5549 Asn Glu Asp Gln Ala Met Met GlnAsp Cys Tyr Ser Lys Ile Val Asp 1790 1795 1800 aag ctg tcc tct gca aacccc acc atg gta tta cag gtt cag atg ctc 5597 Lys Leu Ser Ser Ala Asn ProThr Met Val Leu Gln Val Gln Met Leu 1805 1810 1815 gtg gct gaa ctg cgcagg gtc act gtg ctc tgg gat gag ctc tgg ctg 5645 Val Ala Glu Leu Arg ArgVal Thr Val Leu Trp Asp Glu Leu Trp Leu 1820 1825 1830 1835 gga gtt ttgctg caa caa cac atg tat gtc ctg aga cga att cag cag 5693 Gly Val Leu LeuGln Gln His Met Tyr Val Leu Arg Arg Ile Gln Gln 1840 1845 1850 ctt gaagat gag gtg aag aga gtc cag aac aac aac acc tta cgc aaa 5741 Leu Glu AspGlu Val Lys Arg Val Gln Asn Asn Asn Thr Leu Arg Lys 1855 1860 1865 gaagag aaa att gca atc atg agg gag agg cac aca gct ttg atg aag 5789 Glu GluLys Ile Ala Ile Met Arg Glu Arg His Thr Ala Leu Met Lys 1870 1875 1880ccc atc gta ttt gct ttg gag cat gtg agg agt atc aca gcg gct cct 5837 ProIle Val Phe Ala Leu Glu His Val Arg Ser Ile Thr Ala Ala Pro 1885 18901895 gca gaa aca cct cat gaa aaa tgg ttt cag gat aac tat ggt gat gcc5885 Ala Glu Thr Pro His Glu Lys Trp Phe Gln Asp Asn Tyr Gly Asp Ala1900 1905 1910 1915 att gaa aat gcc cta gaa aaa ctg aag act cca ttg aaccct gca aag 5933 Ile Glu Asn Ala Leu Glu Lys Leu Lys Thr Pro Leu Asn ProAla Lys 1920 1925 1930 cct ggg agc agc tgg att cca ttt aaa gag ata atgcta agt ttg caa 5981 Pro Gly Ser Ser Trp Ile Pro Phe Lys Glu Ile Met LeuSer Leu Gln 1935 1940 1945 cag aga gca cag aaa cgt gca agt tac atc ttgcgt ctt gaa gaa atc 6029 Gln Arg Ala Gln Lys Arg Ala Ser Tyr Ile Leu ArgLeu Glu Glu Ile 1950 1955 1960 agt cca tgg ttg gct gcc atg act aac actgaa att gct ctt cct ggg 6077 Ser Pro Trp Leu Ala Ala Met Thr Asn Thr GluIle Ala Leu Pro Gly 1965 1970 1975 gaa gtc tca gcc aga gac act gtc acaatc cat agt gtg ggc gga acc 6125 Glu Val Ser Ala Arg Asp Thr Val Thr IleHis Ser Val Gly Gly Thr 1980 1985 1990 1995 atc aca atc tta ccg act aaaacc aag cca aag aaa ctt ctc ttt ctt 6173 Ile Thr Ile Leu Pro Thr Lys ThrLys Pro Lys Lys Leu Leu Phe Leu 2000 2005 2010 gga tca gat ggg aag agctat cct tat ctt ttc aaa gga ctg gag gat 6221 Gly Ser Asp Gly Lys Ser TyrPro Tyr Leu Phe Lys Gly Leu Glu Asp 2015 2020 2025 tta cat ctg gat gagaga ata atg cag ttc cta tct att gtg aat acc 6269 Leu His Leu Asp Glu ArgIle Met Gln Phe Leu Ser Ile Val Asn Thr 2030 2035 2040 atg ttt gct acaatt aat cgc caa gaa aca ccc cgg ttc cat gct cga 6317 Met Phe Ala Thr IleAsn Arg Gln Glu Thr Pro Arg Phe His Ala Arg 2045 2050 2055 cac tat tctgta aca cca cta gga aca aga tca gga cta atc cag tgg 6365 His Tyr Ser ValThr Pro Leu Gly Thr Arg Ser Gly Leu Ile Gln Trp 2060 2065 2070 2075 gtagat gga gcc aca ccc tta ttt ggt ctt tac aaa cga tgg caa caa 6413 Val AspGly Ala Thr Pro Leu Phe Gly Leu Tyr Lys Arg Trp Gln Gln 2080 2085 2090cgg gaa gct gcc tta caa gca caa aag gcc caa gat tcc tac caa act 6461 ArgGlu Ala Ala Leu Gln Ala Gln Lys Ala Gln Asp Ser Tyr Gln Thr 2095 21002105 cct cag aat cct gga att gta ccc cgt cct agt gaa ctt tat tac agt6509 Pro Gln Asn Pro Gly Ile Val Pro Arg Pro Ser Glu Leu Tyr Tyr Ser2110 2115 2120 aaa att ggc cct gct ttg aaa aca gtt ggg ctt agc ctg gatgtg tcc 6557 Lys Ile Gly Pro Ala Leu Lys Thr Val Gly Leu Ser Leu Asp ValSer 2125 2130 2135 cgt cgg gat tgg cct ctt cat gta atg aag gca gta ttggaa gag tta 6605 Arg Arg Asp Trp Pro Leu His Val Met Lys Ala Val Leu GluGlu Leu 2140 2145 2150 2155 atg gag gcc aca ccc ccg aat ctc ctt gcc aaagag ctc tgg tca tct 6653 Met Glu Ala Thr Pro Pro Asn Leu Leu Ala Lys GluLeu Trp Ser Ser 2160 2165 2170 tgc aca aca cct gat gaa tgg tgg aga gttacg cag tct tat gca aga 6701 Cys Thr Thr Pro Asp Glu Trp Trp Arg Val ThrGln Ser Tyr Ala Arg 2175 2180 2185 tct act gca gtc atg tct atg gtt ggatac ata att ggc ctt gga gac 6749 Ser Thr Ala Val Met Ser Met Val Gly TyrIle Ile Gly Leu Gly Asp 2190 2195 2200 aga cat ctg gat aat gtt ctt atagat atg acg act gga gaa gtt gtt 6797 Arg His Leu Asp Asn Val Leu Ile AspMet Thr Thr Gly Glu Val Val 2205 2210 2215 cac ata gat tac aat gtt tgcttt gaa aaa ggt aaa agc ctt aga gtt 6845 His Ile Asp Tyr Asn Val Cys PheGlu Lys Gly Lys Ser Leu Arg Val 2220 2225 2230 2235 cct gag aaa gta cctttt cga atg aca caa aac att gaa aca gca ctg 6893 Pro Glu Lys Val Pro PheArg Met Thr Gln Asn Ile Glu Thr Ala Leu 2240 2245 2250 ggt gta act ggagta gaa ggt gta ttt agg ctt tca tgt gag cag gtt 6941 Gly Val Thr Gly ValGlu Gly Val Phe Arg Leu Ser Cys Glu Gln Val 2255 2260 2265 tta cac attatg cgg cgt ggc aga gag acc ctg ctg acg ctg ctg gag 6989 Leu His Ile MetArg Arg Gly Arg Glu Thr Leu Leu Thr Leu Leu Glu 2270 2275 2280 gcc tttgtg tac gac cct ctg gtg gac tgg aca gca gga ggc gag gct 7037 Ala Phe ValTyr Asp Pro Leu Val Asp Trp Thr Ala Gly Gly Glu Ala 2285 2290 2295 gggttt gct ggt gct gtc tat ggt gga ggt ggc cag cag gcc gag agc 7085 Gly PheAla Gly Ala Val Tyr Gly Gly Gly Gly Gln Gln Ala Glu Ser 2300 2305 23102315 aag cag agc aag aga gag atg gag cga gag atc acc cgc agc ctg ttt7133 Lys Gln Ser Lys Arg Glu Met Glu Arg Glu Ile Thr Arg Ser Leu Phe2320 2325 2330 tct tct aga gta gct gag att aag gtg aac tgg ttt aag aataga gat 7181 Ser Ser Arg Val Ala Glu Ile Lys Val Asn Trp Phe Lys Asn ArgAsp 2335 2340 2345 gag atg ctg gtt gtg ctt ccc aag ttg gac ggt agc ttagat gaa tac 7229 Glu Met Leu Val Val Leu Pro Lys Leu Asp Gly Ser Leu AspGlu Tyr 2350 2355 2360 cta agc ttg caa gag caa ctg aca gat gtg gaa aaactg cag ggc aaa 7277 Leu Ser Leu Gln Glu Gln Leu Thr Asp Val Glu Lys LeuGln Gly Lys 2365 2370 2375 cta ctg gag gaa ata gag ttt cta gaa gga gctgaa ggg gtg gat cat 7325 Leu Leu Glu Glu Ile Glu Phe Leu Glu Gly Ala GluGly Val Asp His 2380 2385 2390 2395 cct tct cat act ctg caa cac agg tattct gag cac acc caa cta cag 7373 Pro Ser His Thr Leu Gln His Arg Tyr SerGlu His Thr Gln Leu Gln 2400 2405 2410 act cag caa aga gct gtt cag gaagca atc cag gtg aag ctg aat gaa 7421 Thr Gln Gln Arg Ala Val Gln Glu AlaIle Gln Val Lys Leu Asn Glu 2415 2420 2425 ttt gaa caa tgg ata aca cattat cag gct gca ttc aat aat tta gaa 7469 Phe Glu Gln Trp Ile Thr His TyrGln Ala Ala Phe Asn Asn Leu Glu 2430 2435 2440 gca aca cag ctt gca agcttg ctt caa gag ata agc aca caa atg gac 7517 Ala Thr Gln Leu Ala Ser LeuLeu Gln Glu Ile Ser Thr Gln Met Asp 2445 2450 2455 ctt ggt cct cca agttac gtg cca gca aca gcc ttt ctg cag aat gct 7565 Leu Gly Pro Pro Ser TyrVal Pro Ala Thr Ala Phe Leu Gln Asn Ala 2460 2465 2470 2475 ggt cag gcccac ttg att agc cag tgc gag cag ctg gag ggg gag gtt 7613 Gly Gln Ala HisLeu Ile Ser Gln Cys Glu Gln Leu Glu Gly Glu Val 2480 2485 2490 ggt gctctc ctg cag cag agg cgc tcc gtg ctc cgt ggc tgt ctg gag 7661 Gly Ala LeuLeu Gln Gln Arg Arg Ser Val Leu Arg Gly Cys Leu Glu 2495 2500 2505 caactg cat cac tat gca acc gtg gcc ctg cag tat ccg aag gcc ata 7709 Gln LeuHis His Tyr Ala Thr Val Ala Leu Gln Tyr Pro Lys Ala Ile 2510 2515 2520ttt cag aaa cat cga att gaa cag tgg aag acc tgg atg gaa gag ctc 7757 PheGln Lys His Arg Ile Glu Gln Trp Lys Thr Trp Met Glu Glu Leu 2525 25302535 atc tgt aac acc aca gta gag cgt tgt caa gag ctc tat agg aaa tat7805 Ile Cys Asn Thr Thr Val Glu Arg Cys Gln Glu Leu Tyr Arg Lys Tyr2540 2545 2550 2555 gaa atg caa tat gct ccc cag cca ccc cca aca gtg tgtcag ttc atc 7853 Glu Met Gln Tyr Ala Pro Gln Pro Pro Pro Thr Val Cys GlnPhe Ile 2560 2565 2570 act gcc act gaa atg acc ctg cag cga tac gca gcagac atc aac agc 7901 Thr Ala Thr Glu Met Thr Leu Gln Arg Tyr Ala Ala AspIle Asn Ser 2575 2580 2585 aga ctt att aga caa gtg gaa cgc ttg aaa caggaa gct gtc act gtg 7949 Arg Leu Ile Arg Gln Val Glu Arg Leu Lys Gln GluAla Val Thr Val 2590 2595 2600 cca gtt tgt gaa gat cag ttg aaa gaa attgaa cgt tgc att aaa gtt 7997 Pro Val Cys Glu Asp Gln Leu Lys Glu Ile GluArg Cys Ile Lys Val 2605 2610 2615 ttc ctt cat gag aat gga gaa gaa ggatct ttg agt cta gca agt gtt 8045 Phe Leu His Glu Asn Gly Glu Glu Gly SerLeu Ser Leu Ala Ser Val 2620 2625 2630 2635 att att tct gcc ctt tgt accctt aca agg cgt aac ctg atg atg gaa 8093 Ile Ile Ser Ala Leu Cys Thr LeuThr Arg Arg Asn Leu Met Met Glu 2640 2645 2650 ggt gca gcg tca agt gctgga gaa cag ctg gtt gat ctg act tct cgg 8141 Gly Ala Ala Ser Ser Ala GlyGlu Gln Leu Val Asp Leu Thr Ser Arg 2655 2660 2665 gat gga gcc tgg ttcttg gag gaa ctc tgc agt atg agc gga aac gtc 8189 Asp Gly Ala Trp Phe LeuGlu Glu Leu Cys Ser Met Ser Gly Asn Val 2670 2675 2680 acc tgc ttg gttcag tta ctg aag cag tgc cac ctg gtg cca cag gac 8237 Thr Cys Leu Val GlnLeu Leu Lys Gln Cys His Leu Val Pro Gln Asp 2685 2690 2695 tta gat atcccg aac ccc atg gaa gcg tct gag aca gtt cac tta gcc 8285 Leu Asp Ile ProAsn Pro Met Glu Ala Ser Glu Thr Val His Leu Ala 2700 2705 2710 2715 aatgga gtg tat acc tca ctt cag gaa ttg aat tcg aat ttc cgg caa 8333 Asn GlyVal Tyr Thr Ser Leu Gln Glu Leu Asn Ser Asn Phe Arg Gln 2720 2725 2730atc ata ttt cca gaa gca ctt cga tgt tta atg aaa ggg gaa tac acg 8381 IleIle Phe Pro Glu Ala Leu Arg Cys Leu Met Lys Gly Glu Tyr Thr 2735 27402745 tta gaa agt atg ctg cat gaa ctg gac ggt ctt att gag cag acc acc8429 Leu Glu Ser Met Leu His Glu Leu Asp Gly Leu Ile Glu Gln Thr Thr2750 2755 2760 gat ggc gtt ccc ctg cag act cta gtg gaa tct ctt cag gcctac tta 8477 Asp Gly Val Pro Leu Gln Thr Leu Val Glu Ser Leu Gln Ala TyrLeu 2765 2770 2775 aga aac gca gct atg gga ctg gaa gaa gaa aca cat gctcat tac atc 8525 Arg Asn Ala Ala Met Gly Leu Glu Glu Glu Thr His Ala HisTyr Ile 2780 2785 2790 2795 gat gtt gcc aga cta cta cat gct cag tac ggtgaa tta atc caa ccg 8573 Asp Val Ala Arg Leu Leu His Ala Gln Tyr Gly GluLeu Ile Gln Pro 2800 2805 2810 aga aat ggt tca gtt gat gaa aca ccc aaaatg tca gct ggc cag atg 8621 Arg Asn Gly Ser Val Asp Glu Thr Pro Lys MetSer Ala Gly Gln Met 2815 2820 2825 ctt ttg gta gca ttc gat ggc atg tttgct caa gtt gaa act gct ttc 8669 Leu Leu Val Ala Phe Asp Gly Met Phe AlaGln Val Glu Thr Ala Phe 2830 2835 2840 agc tta tta gtt gaa aag ttg aacaag atg gaa att ccc ata gct tgg 8717 Ser Leu Leu Val Glu Lys Leu Asn LysMet Glu Ile Pro Ile Ala Trp 2845 2850 2855 cga aag att gac atc ata agggaa gcc agg agt act caa gtt aat ttt 8765 Arg Lys Ile Asp Ile Ile Arg GluAla Arg Ser Thr Gln Val Asn Phe 2860 2865 2870 2875 ttt gat gat gat aatcac cgg cag gtg cta gaa gag att ttc ttt cta 8813 Phe Asp Asp Asp Asn HisArg Gln Val Leu Glu Glu Ile Phe Phe Leu 2880 2885 2890 aaa aga cta cagact att aag gag ttc ttc agg ctc tgt ggt acc ttt 8861 Lys Arg Leu Gln ThrIle Lys Glu Phe Phe Arg Leu Cys Gly Thr Phe 2895 2900 2905 tct aaa acattg tca gga tca agt tca ctt gaa gat cag aat act gtg 8909 Ser Lys Thr LeuSer Gly Ser Ser Ser Leu Glu Asp Gln Asn Thr Val 2910 2915 2920 aat gggcct gta cag att gtc aat gtg aaa acc ctt ttt aga aac tct 8957 Asn Gly ProVal Gln Ile Val Asn Val Lys Thr Leu Phe Arg Asn Ser 2925 2930 2935 tgtttc agt gaa gac caa atg gcc aaa cct atc aag gca ttc aca gct 9005 Cys PheSer Glu Asp Gln Met Ala Lys Pro Ile Lys Ala Phe Thr Ala 2940 2945 29502955 gac ttt gtg agg cag ctc ttg ata ggg cta ccc aac caa gcc ctc gga9053 Asp Phe Val Arg Gln Leu Leu Ile Gly Leu Pro Asn Gln Ala Leu Gly2960 2965 2970 ctc aca ctg tgc agt ttt atc agt gct ctg ggt gta gac atcatt gct 9101 Leu Thr Leu Cys Ser Phe Ile Ser Ala Leu Gly Val Asp Ile IleAla 2975 2980 2985 caa gta gag gca aag gac ttt ggt gcc gaa agc aaa gtttct gtt gat 9149 Gln Val Glu Ala Lys Asp Phe Gly Ala Glu Ser Lys Val SerVal Asp 2990 2995 3000 gat ctc tgt aag aaa gcg gtg gaa cat aac atc cagata ggg aag ttc 9197 Asp Leu Cys Lys Lys Ala Val Glu His Asn Ile Gln IleGly Lys Phe 3005 3010 3015 tct cag ctg gtt atg aac agg gca act gtg ttagca agt tct tac gac 9245 Ser Gln Leu Val Met Asn Arg Ala Thr Val Leu AlaSer Ser Tyr Asp 3020 3025 3030 3035 act gcc tgg aag aag cat gac ttg gtgcga agg cta gaa acc agt att 9293 Thr Ala Trp Lys Lys His Asp Leu Val ArgArg Leu Glu Thr Ser Ile 3040 3045 3050 tct tct tgt aag aca agc ctg cagcgg gtt cag ctg cat att gcc atg 9341 Ser Ser Cys Lys Thr Ser Leu Gln ArgVal Gln Leu His Ile Ala Met 3055 3060 3065 ttt cag tgg caa cat gaa gatcta ctt atc aat aga cca caa gcc atg 9389 Phe Gln Trp Gln His Glu Asp LeuLeu Ile Asn Arg Pro Gln Ala Met 3070 3075 3080 tca gtc aca cct ccc ccacgg tct gct atc cta acc agc atg aaa aag 9437 Ser Val Thr Pro Pro Pro ArgSer Ala Ile Leu Thr Ser Met Lys Lys 3085 3090 3095 aag ctg cat acc ctgagc cag att gaa act tct att gcg aca gtt cag 9485 Lys Leu His Thr Leu SerGln Ile Glu Thr Ser Ile Ala Thr Val Gln 3100 3105 3110 3115 gag aag ctagct gca ctt gaa tca agt att gaa cag cga ctc aag tgg 9533 Glu Lys Leu AlaAla Leu Glu Ser Ser Ile Glu Gln Arg Leu Lys Trp 3120 3125 3130 gca ggtggt gcc aac cct gca ttg gcc cct gta cta caa gat ttt gaa 9581 Ala Gly GlyAla Asn Pro Ala Leu Ala Pro Val Leu Gln Asp Phe Glu 3135 3140 3145 gcaacg ata gct gaa aga aga aat ctt gtc ctt aaa gag agc caa aga 9629 Ala ThrIle Ala Glu Arg Arg Asn Leu Val Leu Lys Glu Ser Gln Arg 3150 3155 3160gca agt cag gtc aca ttt ctc tgc agc aat atc att cat ttt gaa agt 9677 AlaSer Gln Val Thr Phe Leu Cys Ser Asn Ile Ile His Phe Glu Ser 3165 31703175 tta cga aca aga act gca gaa gcc tta aac ctg gat gcg gcg tta ttt9725 Leu Arg Thr Arg Thr Ala Glu Ala Leu Asn Leu Asp Ala Ala Leu Phe3180 3185 3190 3195 gaa cta atc aag cga tgt cag cag atg tgt tcg ttt gcatca cag ttt 9773 Glu Leu Ile Lys Arg Cys Gln Gln Met Cys Ser Phe Ala SerGln Phe 3200 3205 3210 aac agt tca gtg tct gag tta gag ctt cgt tta ttacag aga gtg gac 9821 Asn Ser Ser Val Ser Glu Leu Glu Leu Arg Leu Leu GlnArg Val Asp 3215 3220 3225 act ggt ctt gaa cat cct att ggc agc tct gaatgg ctt ttg tca gca 9869 Thr Gly Leu Glu His Pro Ile Gly Ser Ser Glu TrpLeu Leu Ser Ala 3230 3235 3240 cac aaa cag ttg acc cag gat atg tct actcag agg gca att cag aca 9917 His Lys Gln Leu Thr Gln Asp Met Ser Thr GlnArg Ala Ile Gln Thr 3245 3250 3255 gag aaa gag cag cag ata gaa acg gtctgt gaa aca att cag aat ctg 9965 Glu Lys Glu Gln Gln Ile Glu Thr Val CysGlu Thr Ile Gln Asn Leu 3260 3265 3270 3275 gtt gat aat ata aag act gtgctc act ggt cat aac cga cag ctt gga 10013 Val Asp Asn Ile Lys Thr ValLeu Thr Gly His Asn Arg Gln Leu Gly 3280 3285 3290 gat gtc aaa cat ctcttg aaa gct atg gct aag gat gaa gaa gct gct 10061 Asp Val Lys His LeuLeu Lys Ala Met Ala Lys Asp Glu Glu Ala Ala 3295 3300 3305 ctg gca gatggt gaa gat gtt ccc tat gag aac agt gtt agg cag ttt 10109 Leu Ala AspGly Glu Asp Val Pro Tyr Glu Asn Ser Val Arg Gln Phe 3310 3315 3320 ttgggt gaa tat aaa tca tgg caa gac aac att caa aca gtt cta ttt 10157 LeuGly Glu Tyr Lys Ser Trp Gln Asp Asn Ile Gln Thr Val Leu Phe 3325 33303335 aca tta gtc cag gct atg ggt cag gtt cga agt caa gaa cac gtt gaa10205 Thr Leu Val Gln Ala Met Gly Gln Val Arg Ser Gln Glu His Val Glu3340 3345 3350 3355 atg ctc cag gaa atc act ccc acc ttg aaa gaa ctg aaaaca caa agt 10253 Met Leu Gln Glu Ile Thr Pro Thr Leu Lys Glu Leu LysThr Gln Ser 3360 3365 3370 cag agt atc tat aat aat tta gtg agt ttt gcatca ccc tta gtc acc 10301 Gln Ser Ile Tyr Asn Asn Leu Val Ser Phe AlaSer Pro Leu Val Thr 3375 3380 3385 gat gca aca aat gaa tgt tcg agt ccaacg tca tct gct act tat cag 10349 Asp Ala Thr Asn Glu Cys Ser Ser ProThr Ser Ser Ala Thr Tyr Gln 3390 3395 3400 cca tcc ttc gct gca gca gtccgg agt aac act ggc cag aag act cag 10397 Pro Ser Phe Ala Ala Ala ValArg Ser Asn Thr Gly Gln Lys Thr Gln 3405 3410 3415 cct gat gtc atg tcacag aat gct aga aag ctg atc cag aaa aat ctt 10445 Pro Asp Val Met SerGln Asn Ala Arg Lys Leu Ile Gln Lys Asn Leu 3420 3425 3430 3435 gct acatca gct gat act cca cca agc acc gtt cca gga act ggc aag 10493 Ala ThrSer Ala Asp Thr Pro Pro Ser Thr Val Pro Gly Thr Gly Lys 3440 3445 3450agt gtt gct tgt agt cct aaa aag gca gtc aga gac cct aaa act ggg 10541Ser Val Ala Cys Ser Pro Lys Lys Ala Val Arg Asp Pro Lys Thr Gly 34553460 3465 aaa gcg gtg caa gag aga aac tcc tat gca gtg agt gtg tgg aagaga 10589 Lys Ala Val Gln Glu Arg Asn Ser Tyr Ala Val Ser Val Trp LysArg 3470 3475 3480 gtg aaa gcc aag tta gag ggc cga gat gtt gat ccg aatagg agg atg 10637 Val Lys Ala Lys Leu Glu Gly Arg Asp Val Asp Pro AsnArg Arg Met 3485 3490 3495 tca gtt gct gaa cag gtt gac tat gtc att aaggaa gca act aat cta 10685 Ser Val Ala Glu Gln Val Asp Tyr Val Ile LysGlu Ala Thr Asn Leu 3500 3505 3510 3515 gat aac ttg gct cag ctg tat gaaggt tgg aca gcc tgg gtg tga 10730 Asp Asn Leu Ala Gln Leu Tyr Glu GlyTrp Thr Ala Trp Val * 3520 3525 atggcaagac agtagatgag tctggttaagcgaggtcaga catccaccag aatcaactca 10790 gcctcaggca tccaaagcca caccacagtcggtggtgatg caactggggg cttactctga 10850 ggaaacctag gaaatctcgg tgcactaggaagtgaatccc gcaggacagc tgcactcagg 10910 gatacgccca acaccatggc ctgcaaccccagggtcaagg gtgaaggaaa gcaaagctca 10970 ccgcctgaac acggagattg tctttctgccacagaacagc agcagacgtg tcgggaggtt 11030 agctgcggaa agaaatcggg atgccgcggagcacagagtg atttggaact ccattccacc 11090 tgaccctgtg tgtacaatcc aggaaaaaaacaaaccccac tcagaaacag agaaaactgg 11150 ggtcgcgaag aaatcacagc caaggaagatttgatgcatt cagattctcg tgtaacactt 11210 gttgcttggc aacagtactg gttgggttgaccagtaagta gaaaaaggct aaaggctatg 11270 cgatatgaat ttcagaaatg gactgaaaatggagagctat gtaacagata cactacagta 11330 gaagaactta cttctgaaat gaagggaaaaaaaccacccc atcgttccct actcctcccc 11390 accacttacc cgttccccct ttacctaatctagtagatta gccatctttc aaattcactt 11450 ttatttcagt ccttatattt catatacttccgtctcgatg ctgttaacaa cttctgataa 11510 catggaaaat tcaaggattg tttaaaggtctgatgatcac acacaaaatg taattccggt 11570 tatttaagtc atttctgtga ttctatcatgtacagtttcc agaattgtca ctgtgcattc 11630 aaaagtaatg aatctaacag acatttgatttaatgtacac tcccttttgc ttatagtgtg 11690 catttttttt ggaggtcatt caaattttccctcttctgtg atagctgtag tttctttcat 11750 agaaagtagc taatccagtg taatcttttacctttttaaa aaccaagata gagtatctat 11810 tagagtttta cattgttgat gatagattaacaataaagtg atgttctggt ggaggtagac 11870 tgaaattttt ttaattcatg tttttcatttgatactttta atttacactt agtaaattaa 11930 aagttgttta atttacttgg cattttaggacatgtacatg aaacagtgaa aatgagatcc 11990 accaacatct tttattaagt tcagttattagtctgtgaag tgctttactt tttgcacaat 12050 tttaatagct tgctattcag taatacattatagtgaattc atgatcaagg tttccttaaa 12110 tttagcattg catttcagta ctgactgtgtaagctaaatt gctgatccaa aataaaaacc 12170 cagactagaa tagggttctt aaaatcaagtatcaatacaa aatagaacac aattaaaatc 12230 ttaattgttg gctgggcaca gtggctcacgcctgtaatcc cagcactttg ggaggccgag 12290 gcgggcggat catgaggtta ggagagcgagaccatcctgg ctaacacggt gaaaccccgt 12350 ctttactaaa atacaaaaaa aattagccgggtgtggtggc gggcgcctgt agtcccagct 12410 actcgggagg ctgaggcagg agaatggcgtgaacccagga ggcggagctt gcagtgagcc 12470 gagattgtgc cactgcactc cagcctgggcaacagagcta gactctgtgt caaaaataaa 12530 tgactagat 12539 8 3529 PRT Homosapiens 8 Met Arg Lys Ser Gln Glu Arg Ser Met Ser Tyr Ser Asp Glu SerArg 1 5 10 15 Leu Ser Asn Leu Leu Arg Arg Ile Thr Arg Glu Asp Asp ArgAsp Arg 20 25 30 Arg Leu Ala Thr Val Lys Gln Leu Lys Glu Phe Ile Gln GlnPro Glu 35 40 45 Asn Lys Leu Val Leu Val Lys Gln Leu Asp Asn Ile Leu AlaAla Val 50 55 60 His Asp Val Leu Asn Glu Ser Ser Lys Leu Leu Gln Glu LeuArg Gln 65 70 75 80 Glu Gly Ala Cys Cys Leu Gly Leu Leu Cys Ala Ser LeuSer Tyr Glu 85 90 95 Ala Glu Lys Ile Phe Lys Trp Ile Phe Ser Lys Phe SerSer Ser Ala 100 105 110 Lys Asp Glu Val Lys Leu Leu Tyr Leu Cys Ala ThrTyr Lys Ala Leu 115 120 125 Glu Thr Val Gly Glu Lys Lys Ala Phe Ser SerVal Met Gln Leu Val 130 135 140 Met Thr Ser Leu Gln Ser Ile Leu Glu AsnVal Asp Thr Pro Glu Leu 145 150 155 160 Leu Cys Lys Cys Val Lys Cys IleLeu Leu Val Ala Arg Cys Tyr Pro 165 170 175 His Ile Phe Ser Thr Asn PheArg Asp Thr Val Asp Ile Leu Val Gly 180 185 190 Trp His Ile Asp His ThrGln Lys Pro Ser Leu Thr Gln Gln Val Ser 195 200 205 Gly Trp Leu Gln SerLeu Glu Pro Phe Trp Val Ala Asp Leu Ala Phe 210 215 220 Ser Thr Thr LeuLeu Gly Gln Phe Leu Glu Asp Met Glu Ala Tyr Ala 225 230 235 240 Glu AspLeu Ser His Val Ala Ser Gly Glu Ser Val Asp Glu Asp Val 245 250 255 ProPro Pro Ser Val Ser Leu Pro Lys Leu Ala Ala Leu Leu Arg Val 260 265 270Phe Ser Thr Val Val Arg Ser Ile Gly Glu Arg Phe Ser Pro Ile Arg 275 280285 Gly Pro Pro Ile Thr Glu Ala Tyr Val Thr Asp Val Leu Tyr Arg Val 290295 300 Met Arg Cys Val Thr Ala Ala Asn Gln Val Phe Phe Ser Glu Ala Val305 310 315 320 Leu Thr Ala Ala Asn Glu Cys Val Gly Val Leu Leu Gly SerLeu Asp 325 330 335 Pro Ser Met Thr Ile His Cys Asp Met Val Ile Thr TyrGly Leu Asp 340 345 350 Gln Leu Glu Asn Cys Gln Thr Cys Gly Thr Asp TyrIle Ile Ser Val 355 360 365 Leu Asn Leu Leu Thr Leu Ile Val Glu Gln IleAsn Thr Lys Leu Pro 370 375 380 Ser Ser Phe Val Glu Lys Leu Phe Ile ProSer Ser Lys Leu Leu Phe 385 390 395 400 Leu Arg Tyr His Lys Glu Lys GluVal Val Ala Val Ala His Ala Val 405 410 415 Tyr Gln Ala Val Leu Ser LeuLys Asn Ile Pro Val Leu Glu Thr Ala 420 425 430 Tyr Lys Leu Ile Leu GlyGlu Met Thr Cys Ala Leu Asn Asn Leu Leu 435 440 445 His Ser Leu Gln LeuPro Glu Ala Cys Ser Glu Ile Lys His Glu Ala 450 455 460 Phe Lys Asn HisVal Phe Asn Val Asp Asn Ala Lys Phe Val Val Lys 465 470 475 480 Phe AspLeu Ser Ala Leu Thr Thr Ile Gly Asn Ala Lys Asn Ser Leu 485 490 495 IleGly Met Trp Ala Leu Ser Pro Thr Val Phe Ala Leu Leu Ser Lys 500 505 510Asn Leu Met Ile Val His Ser Asp Leu Ala Val His Phe Pro Ala Ile 515 520525 Gln Tyr Ala Val Leu Tyr Thr Leu Tyr Ser His Cys Thr Arg His Asp 530535 540 His Phe Ile Ser Ser Ser Leu Ser Ser Ala Ser Pro Ser Leu Phe Asp545 550 555 560 Gly Ala Val Ile Ser Thr Val Thr Thr Ala Thr Lys Lys HisPhe Ser 565 570 575 Ile Ile Leu Asn Leu Leu Gly Ile Leu Leu Lys Lys AspAsn Leu Asn 580 585 590 Gln Asp Thr Arg Lys Leu Leu Met Thr Trp Ala LeuGlu Ala Ala Val 595 600 605 Leu Met Arg Lys Ser Glu Thr Tyr Ala Pro LeuPhe Ser Leu Pro Ser 610 615 620 Phe His Lys Phe Cys Lys Gly Leu Leu AlaAsn Thr Leu Val Glu Asp 625 630 635 640 Val Asn Ile Cys Leu Gln Ala CysSer Ser Leu His Ala Leu Ser Ser 645 650 655 Ser Leu Pro Asp Asp Leu LeuGln Arg Cys Val Asp Val Cys Arg Val 660 665 670 Gln Leu Val His Ser GlyThr Arg Ile Arg Gln Ala Phe Gly Lys Leu 675 680 685 Leu Lys Ser Ile ProLeu Asp Val Val Leu Ser Asn Asn Asn His Thr 690 695 700 Glu Ile Gln GluIle Ser Leu Ala Leu Arg Ser His Met Ser Lys Ala 705 710 715 720 Pro SerAsn Thr Phe His Pro Gln Asp Phe Ser Asp Val Ile Ser Phe 725 730 735 IleLeu Tyr Gly Asn Ser His Arg Thr Gly Lys Asp Asn Trp Leu Glu 740 745 750Arg Leu Phe Tyr Ser Cys Gln Arg Leu Asp Lys Arg Asp Gln Ser Thr 755 760765 Ile Pro Arg Asn Leu Leu Lys Thr Asp Ala Val Leu Trp Gln Trp Ala 770775 780 Ile Trp Glu Ala Ala Gln Phe Thr Val Leu Ser Lys Leu Arg Thr Pro785 790 795 800 Leu Gly Arg Ala Gln Asp Thr Phe Gln Thr Ile Glu Gly IleIle Arg 805 810 815 Ser Leu Ala Ala His Thr Leu Asn Pro Asp Gln Asp ValSer Gln Trp 820 825 830 Thr Thr Ala Asp Asn Asp Glu Gly His Gly Asn AsnGln Leu Arg Leu 835 840 845 Val Leu Leu Leu Gln Tyr Leu Glu Asn Leu GluLys Leu Met Tyr Asn 850 855 860 Ala Tyr Glu Gly Cys Ala Asn Ala Leu ThrSer Pro Pro Lys Val Ile 865 870 875 880 Arg Thr Phe Phe Tyr Thr Asn ArgGln Thr Cys Gln Asp Trp Leu Thr 885 890 895 Arg Ile Arg Leu Ser Ile MetArg Val Gly Leu Leu Ala Gly Gln Pro 900 905 910 Ala Val Thr Val Arg HisGly Phe Asp Leu Leu Thr Glu Met Lys Thr 915 920 925 Thr Ser Leu Ser GlnGly Asn Glu Leu Glu Val Thr Ile Met Met Val 930 935 940 Val Glu Ala LeuCys Glu Leu His Cys Pro Glu Ala Ile Gln Gly Ile 945 950 955 960 Ala ValTrp Ser Ser Ser Ile Val Gly Lys Asn Leu Leu Trp Ile Asn 965 970 975 SerVal Ala Gln Gln Ala Glu Gly Arg Phe Glu Lys Ala Ser Val Glu 980 985 990Tyr Gln Glu His Leu Cys Ala Met Thr Gly Val Asp Cys Cys Ile Ser 995 10001005 Ser Phe Asp Lys Ser Val Leu Thr Leu Ala Asn Ala Gly Arg Asn Ser1010 1015 1020 Ala Ser Pro Lys His Ser Leu Asn Gly Glu Ser Arg Lys ThrVal Leu 1025 1030 1035 1040 Ser Lys Pro Thr Asp Ser Ser Pro Glu Val IleAsn Tyr Leu Gly Asn 1045 1050 1055 Lys Ala Cys Glu Phe Tyr Ile Ser IleAla Asp Trp Ala Ala Val Gln 1060 1065 1070 Glu Trp Gln Asn Ala Ile HisAsp Leu Lys Lys Ser Thr Ser Ser Thr 1075 1080 1085 Ser Leu Asn Leu LysAla Asp Phe Asn Tyr Ile Lys Ser Leu Ser Ser 1090 1095 1100 Phe Glu SerGly Lys Phe Val Glu Cys Thr Glu Gln Leu Glu Leu Leu 1105 1110 1115 1120Pro Gly Glu Asn Ile Asn Leu Leu Ala Gly Gly Ser Lys Glu Lys Ile 11251130 1135 Asp Met Lys Lys Leu Leu Pro Asn Met Leu Ser Pro Asp Pro ArgGlu 1140 1145 1150 Leu Gln Lys Ser Ile Glu Val Gln Leu Leu Arg Ser SerVal Cys Leu 1155 1160 1165 Ala Thr Ala Leu Asn Pro Ile Glu Gln Asp GlnLys Trp Gln Ser Ile 1170 1175 1180 Thr Glu Asn Val Val Lys Tyr Leu LysGln Thr Ser Arg Ile Ala Ile 1185 1190 1195 1200 Gly Pro Leu Arg Leu SerThr Leu Thr Val Ser Gln Ser Leu Pro Val 1205 1210 1215 Leu Ser Thr LeuGln Leu Tyr Cys Ser Ser Ala Leu Glu Asn Thr Val 1220 1225 1230 Ser AsnArg Leu Ser Thr Glu Asp Cys Leu Ile Pro Leu Phe Ser Glu 1235 1240 1245Ala Leu Arg Ser Cys Lys Gln His Asp Val Arg Pro Trp Met Gln Ala 12501255 1260 Leu Arg Tyr Thr Met Tyr Gln Asn Gln Leu Leu Glu Lys Ile LysGlu 1265 1270 1275 1280 Gln Thr Val Pro Ile Arg Ser His Leu Met Glu LeuGly Leu Thr Ala 1285 1290 1295 Ala Lys Phe Ala Arg Lys Arg Gly Asn ValSer Leu Ala Thr Arg Leu 1300 1305 1310 Leu Ala Gln Cys Ser Glu Val GlnLeu Gly Lys Thr Thr Thr Ala Gln 1315 1320 1325 Asp Leu Val Gln His PheLys Lys Leu Ser Thr Gln Gly Gln Val Asp 1330 1335 1340 Glu Lys Trp GlyPro Glu Leu Asp Ile Glu Lys Thr Lys Leu Leu Tyr 1345 1350 1355 1360 ThrAla Gly Gln Ser Thr His Ala Met Glu Met Leu Ser Ser Cys Ala 1365 13701375 Ile Ser Phe Cys Lys Ser Val Lys Ala Glu Tyr Ala Val Ala Lys Ser1380 1385 1390 Ile Leu Thr Leu Ala Lys Trp Ile Gln Ala Glu Trp Lys GluIle Ser 1395 1400 1405 Gly Gln Leu Lys Gln Val Tyr Arg Ala Gln His GlnGln Asn Phe Thr 1410 1415 1420 Gly Leu Ser Thr Leu Ser Lys Asn Ile LeuThr Leu Ile Glu Leu Pro 1425 1430 1435 1440 Ser Val Asn Thr Met Glu GluGlu Tyr Pro Arg Ile Glu Ser Glu Ser 1445 1450 1455 Thr Val His Ile GlyVal Gly Glu Pro Asp Phe Ile Leu Gly Gln Leu 1460 1465 1470 Tyr His LeuSer Ser Val Gln Ala Pro Glu Val Ala Lys Ser Trp Ala 1475 1480 1485 AlaLeu Ala Ser Trp Ala Tyr Arg Trp Gly Arg Lys Val Val Asp Asn 1490 14951500 Ala Ser Gln Gly Glu Gly Val Arg Leu Leu Pro Arg Glu Lys Ser Glu1505 1510 1515 1520 Val Gln Asn Leu Leu Pro Asp Thr Ile Thr Glu Glu GluLys Glu Arg 1525 1530 1535 Ile Tyr Gly Ile Leu Gly Gln Ala Val Cys ArgPro Ala Gly Ile Gln 1540 1545 1550 Asp Glu Asp Ile Thr Leu Gln Ile ThrGlu Ser Glu Asp Asn Glu Glu 1555 1560 1565 Asp Asp Met Val Asp Val IleTrp Arg Gln Leu Ile Ser Ser Cys Pro 1570 1575 1580 Trp Leu Ser Glu LeuAsp Glu Ser Ala Thr Glu Gly Val Ile Lys Val 1585 1590 1595 1600 Trp ArgLys Val Val Asp Arg Ile Phe Ser Leu Tyr Lys Leu Ser Cys 1605 1610 1615Ser Ala Tyr Phe Thr Phe Leu Lys Leu Asn Ala Gly Gln Ile Pro Leu 16201625 1630 Asp Glu Asp Asp Pro Arg Leu His Leu Ser His Arg Val Glu GlnSer 1635 1640 1645 Thr Asp Asp Met Ile Val Met Ala Thr Leu Arg Leu LeuArg Leu Leu 1650 1655 1660 Val Lys His Ala Gly Glu Leu Arg Gln Tyr LeuGlu His Gly Leu Glu 1665 1670 1675 1680 Thr Thr Pro Thr Ala Pro Trp ArgGly Ile Ile Pro Gln Leu Phe Ser 1685 1690 1695 Arg Leu Asn His Pro GluVal Tyr Val Arg Gln Ser Ile Cys Asn Leu 1700 1705 1710 Leu Cys Arg ValAla Gln Asp Ser Pro His Leu Ile Leu Tyr Pro Ala 1715 1720 1725 Ile ValGly Thr Ile Ser Leu Ser Ser Glu Ser Gln Ala Ser Gly Asn 1730 1735 1740Lys Phe Ser Thr Ala Ile Pro Thr Leu Leu Gly Asn Ile Gln Gly Glu 17451750 1755 1760 Glu Leu Leu Val Ser Glu Cys Glu Gly Gly Ser Pro Pro AlaSer Gln 1765 1770 1775 Asp Ser Asn Lys Asp Glu Pro Lys Ser Gly Leu AsnGlu Asp Gln Ala 1780 1785 1790 Met Met Gln Asp Cys Tyr Ser Lys Ile ValAsp Lys Leu Ser Ser Ala 1795 1800 1805 Asn Pro Thr Met Val Leu Gln ValGln Met Leu Val Ala Glu Leu Arg 1810 1815 1820 Arg Val Thr Val Leu TrpAsp Glu Leu Trp Leu Gly Val Leu Leu Gln 1825 1830 1835 1840 Gln His MetTyr Val Leu Arg Arg Ile Gln Gln Leu Glu Asp Glu Val 1845 1850 1855 LysArg Val Gln Asn Asn Asn Thr Leu Arg Lys Glu Glu Lys Ile Ala 1860 18651870 Ile Met Arg Glu Arg His Thr Ala Leu Met Lys Pro Ile Val Phe Ala1875 1880 1885 Leu Glu His Val Arg Ser Ile Thr Ala Ala Pro Ala Glu ThrPro His 1890 1895 1900 Glu Lys Trp Phe Gln Asp Asn Tyr Gly Asp Ala IleGlu Asn Ala Leu 1905 1910 1915 1920 Glu Lys Leu Lys Thr Pro Leu Asn ProAla Lys Pro Gly Ser Ser Trp 1925 1930 1935 Ile Pro Phe Lys Glu Ile MetLeu Ser Leu Gln Gln Arg Ala Gln Lys 1940 1945 1950 Arg Ala Ser Tyr IleLeu Arg Leu Glu Glu Ile Ser Pro Trp Leu Ala 1955 1960 1965 Ala Met ThrAsn Thr Glu Ile Ala Leu Pro Gly Glu Val Ser Ala Arg 1970 1975 1980 AspThr Val Thr Ile His Ser Val Gly Gly Thr Ile Thr Ile Leu Pro 1985 19901995 2000 Thr Lys Thr Lys Pro Lys Lys Leu Leu Phe Leu Gly Ser Asp GlyLys 2005 2010 2015 Ser Tyr Pro Tyr Leu Phe Lys Gly Leu Glu Asp Leu HisLeu Asp Glu 2020 2025 2030 Arg Ile Met Gln Phe Leu Ser Ile Val Asn ThrMet Phe Ala Thr Ile 2035 2040 2045 Asn Arg Gln Glu Thr Pro Arg Phe HisAla Arg His Tyr Ser Val Thr 2050 2055 2060 Pro Leu Gly Thr Arg Ser GlyLeu Ile Gln Trp Val Asp Gly Ala Thr 2065 2070 2075 2080 Pro Leu Phe GlyLeu Tyr Lys Arg Trp Gln Gln Arg Glu Ala Ala Leu 2085 2090 2095 Gln AlaGln Lys Ala Gln Asp Ser Tyr Gln Thr Pro Gln Asn Pro Gly 2100 2105 2110Ile Val Pro Arg Pro Ser Glu Leu Tyr Tyr Ser Lys Ile Gly Pro Ala 21152120 2125 Leu Lys Thr Val Gly Leu Ser Leu Asp Val Ser Arg Arg Asp TrpPro 2130 2135 2140 Leu His Val Met Lys Ala Val Leu Glu Glu Leu Met GluAla Thr Pro 2145 2150 2155 2160 Pro Asn Leu Leu Ala Lys Glu Leu Trp SerSer Cys Thr Thr Pro Asp 2165 2170 2175 Glu Trp Trp Arg Val Thr Gln SerTyr Ala Arg Ser Thr Ala Val Met 2180 2185 2190 Ser Met Val Gly Tyr IleIle Gly Leu Gly Asp Arg His Leu Asp Asn 2195 2200 2205 Val Leu Ile AspMet Thr Thr Gly Glu Val Val His Ile Asp Tyr Asn 2210 2215 2220 Val CysPhe Glu Lys Gly Lys Ser Leu Arg Val Pro Glu Lys Val Pro 2225 2230 22352240 Phe Arg Met Thr Gln Asn Ile Glu Thr Ala Leu Gly Val Thr Gly Val2245 2250 2255 Glu Gly Val Phe Arg Leu Ser Cys Glu Gln Val Leu His IleMet Arg 2260 2265 2270 Arg Gly Arg Glu Thr Leu Leu Thr Leu Leu Glu AlaPhe Val Tyr Asp 2275 2280 2285 Pro Leu Val Asp Trp Thr Ala Gly Gly GluAla Gly Phe Ala Gly Ala 2290 2295 2300 Val Tyr Gly Gly Gly Gly Gln GlnAla Glu Ser Lys Gln Ser Lys Arg 2305 2310 2315 2320 Glu Met Glu Arg GluIle Thr Arg Ser Leu Phe Ser Ser Arg Val Ala 2325 2330 2335 Glu Ile LysVal Asn Trp Phe Lys Asn Arg Asp Glu Met Leu Val Val 2340 2345 2350 LeuPro Lys Leu Asp Gly Ser Leu Asp Glu Tyr Leu Ser Leu Gln Glu 2355 23602365 Gln Leu Thr Asp Val Glu Lys Leu Gln Gly Lys Leu Leu Glu Glu Ile2370 2375 2380 Glu Phe Leu Glu Gly Ala Glu Gly Val Asp His Pro Ser HisThr Leu 2385 2390 2395 2400 Gln His Arg Tyr Ser Glu His Thr Gln Leu GlnThr Gln Gln Arg Ala 2405 2410 2415 Val Gln Glu Ala Ile Gln Val Lys LeuAsn Glu Phe Glu Gln Trp Ile 2420 2425 2430 Thr His Tyr Gln Ala Ala PheAsn Asn Leu Glu Ala Thr Gln Leu Ala 2435 2440 2445 Ser Leu Leu Gln GluIle Ser Thr Gln Met Asp Leu Gly Pro Pro Ser 2450 2455 2460 Tyr Val ProAla Thr Ala Phe Leu Gln Asn Ala Gly Gln Ala His Leu 2465 2470 2475 2480Ile Ser Gln Cys Glu Gln Leu Glu Gly Glu Val Gly Ala Leu Leu Gln 24852490 2495 Gln Arg Arg Ser Val Leu Arg Gly Cys Leu Glu Gln Leu His HisTyr 2500 2505 2510 Ala Thr Val Ala Leu Gln Tyr Pro Lys Ala Ile Phe GlnLys His Arg 2515 2520 2525 Ile Glu Gln Trp Lys Thr Trp Met Glu Glu LeuIle Cys Asn Thr Thr 2530 2535 2540 Val Glu Arg Cys Gln Glu Leu Tyr ArgLys Tyr Glu Met Gln Tyr Ala 2545 2550 2555 2560 Pro Gln Pro Pro Pro ThrVal Cys Gln Phe Ile Thr Ala Thr Glu Met 2565 2570 2575 Thr Leu Gln ArgTyr Ala Ala Asp Ile Asn Ser Arg Leu Ile Arg Gln 2580 2585 2590 Val GluArg Leu Lys Gln Glu Ala Val Thr Val Pro Val Cys Glu Asp 2595 2600 2605Gln Leu Lys Glu Ile Glu Arg Cys Ile Lys Val Phe Leu His Glu Asn 26102615 2620 Gly Glu Glu Gly Ser Leu Ser Leu Ala Ser Val Ile Ile Ser AlaLeu 2625 2630 2635 2640 Cys Thr Leu Thr Arg Arg Asn Leu Met Met Glu GlyAla Ala Ser Ser 2645 2650 2655 Ala Gly Glu Gln Leu Val Asp Leu Thr SerArg Asp Gly Ala Trp Phe 2660 2665 2670 Leu Glu Glu Leu Cys Ser Met SerGly Asn Val Thr Cys Leu Val Gln 2675 2680 2685 Leu Leu Lys Gln Cys HisLeu Val Pro Gln Asp Leu Asp Ile Pro Asn 2690 2695 2700 Pro Met Glu AlaSer Glu Thr Val His Leu Ala Asn Gly Val Tyr Thr 2705 2710 2715 2720 SerLeu Gln Glu Leu Asn Ser Asn Phe Arg Gln Ile Ile Phe Pro Glu 2725 27302735 Ala Leu Arg Cys Leu Met Lys Gly Glu Tyr Thr Leu Glu Ser Met Leu2740 2745 2750 His Glu Leu Asp Gly Leu Ile Glu Gln Thr Thr Asp Gly ValPro Leu 2755 2760 2765 Gln Thr Leu Val Glu Ser Leu Gln Ala Tyr Leu ArgAsn Ala Ala Met 2770 2775 2780 Gly Leu Glu Glu Glu Thr His Ala His TyrIle Asp Val Ala Arg Leu 2785 2790 2795 2800 Leu His Ala Gln Tyr Gly GluLeu Ile Gln Pro Arg Asn Gly Ser Val 2805 2810 2815 Asp Glu Thr Pro LysMet Ser Ala Gly Gln Met Leu Leu Val Ala Phe 2820 2825 2830 Asp Gly MetPhe Ala Gln Val Glu Thr Ala Phe Ser Leu Leu Val Glu 2835 2840 2845 LysLeu Asn Lys Met Glu Ile Pro Ile Ala Trp Arg Lys Ile Asp Ile 2850 28552860 Ile Arg Glu Ala Arg Ser Thr Gln Val Asn Phe Phe Asp Asp Asp Asn2865 2870 2875 2880 His Arg Gln Val Leu Glu Glu Ile Phe Phe Leu Lys ArgLeu Gln Thr 2885 2890 2895 Ile Lys Glu Phe Phe Arg Leu Cys Gly Thr PheSer Lys Thr Leu Ser 2900 2905 2910 Gly Ser Ser Ser Leu Glu Asp Gln AsnThr Val Asn Gly Pro Val Gln 2915 2920 2925 Ile Val Asn Val Lys Thr LeuPhe Arg Asn Ser Cys Phe Ser Glu Asp 2930 2935 2940 Gln Met Ala Lys ProIle Lys Ala Phe Thr Ala Asp Phe Val Arg Gln 2945 2950 2955 2960 Leu LeuIle Gly Leu Pro Asn Gln Ala Leu Gly Leu Thr Leu Cys Ser 2965 2970 2975Phe Ile Ser Ala Leu Gly Val Asp Ile Ile Ala Gln Val Glu Ala Lys 29802985 2990 Asp Phe Gly Ala Glu Ser Lys Val Ser Val Asp Asp Leu Cys LysLys 2995 3000 3005 Ala Val Glu His Asn Ile Gln Ile Gly Lys Phe Ser GlnLeu Val Met 3010 3015 3020 Asn Arg Ala Thr Val Leu Ala Ser Ser Tyr AspThr Ala Trp Lys Lys 3025 3030 3035 3040 His Asp Leu Val Arg Arg Leu GluThr Ser Ile Ser Ser Cys Lys Thr 3045 3050 3055 Ser Leu Gln Arg Val GlnLeu His Ile Ala Met Phe Gln Trp Gln His 3060 3065 3070 Glu Asp Leu LeuIle Asn Arg Pro Gln Ala Met Ser Val Thr Pro Pro 3075 3080 3085 Pro ArgSer Ala Ile Leu Thr Ser Met Lys Lys Lys Leu His Thr Leu 3090 3095 3100Ser Gln Ile Glu Thr Ser Ile Ala Thr Val Gln Glu Lys Leu Ala Ala 31053110 3115 3120 Leu Glu Ser Ser Ile Glu Gln Arg Leu Lys Trp Ala Gly GlyAla Asn 3125 3130 3135 Pro Ala Leu Ala Pro Val Leu Gln Asp Phe Glu AlaThr Ile Ala Glu 3140 3145 3150 Arg Arg Asn Leu Val Leu Lys Glu Ser GlnArg Ala Ser Gln Val Thr 3155 3160 3165 Phe Leu Cys Ser Asn Ile Ile HisPhe Glu Ser Leu Arg Thr Arg Thr 3170 3175 3180 Ala Glu Ala Leu Asn LeuAsp Ala Ala Leu Phe Glu Leu Ile Lys Arg 3185 3190 3195 3200 Cys Gln GlnMet Cys Ser Phe Ala Ser Gln Phe Asn Ser Ser Val Ser 3205 3210 3215 GluLeu Glu Leu Arg Leu Leu Gln Arg Val Asp Thr Gly Leu Glu His 3220 32253230 Pro Ile Gly Ser Ser Glu Trp Leu Leu Ser Ala His Lys Gln Leu Thr3235 3240 3245 Gln Asp Met Ser Thr Gln Arg Ala Ile Gln Thr Glu Lys GluGln Gln 3250 3255 3260 Ile Glu Thr Val Cys Glu Thr Ile Gln Asn Leu ValAsp Asn Ile Lys 3265 3270 3275 3280 Thr Val Leu Thr Gly His Asn Arg GlnLeu Gly Asp Val Lys His Leu 3285 3290 3295 Leu Lys Ala Met Ala Lys AspGlu Glu Ala Ala Leu Ala Asp Gly Glu 3300 3305 3310 Asp Val Pro Tyr GluAsn Ser Val Arg Gln Phe Leu Gly Glu Tyr Lys 3315 3320 3325 Ser Trp GlnAsp Asn Ile Gln Thr Val Leu Phe Thr Leu Val Gln Ala 3330 3335 3340 MetGly Gln Val Arg Ser Gln Glu His Val Glu Met Leu Gln Glu Ile 3345 33503355 3360 Thr Pro Thr Leu Lys Glu Leu Lys Thr Gln Ser Gln Ser Ile TyrAsn 3365 3370 3375 Asn Leu Val Ser Phe Ala Ser Pro Leu Val Thr Asp AlaThr Asn Glu 3380 3385 3390 Cys Ser Ser Pro Thr Ser Ser Ala Thr Tyr GlnPro Ser Phe Ala Ala 3395 3400 3405 Ala Val Arg Ser Asn Thr Gly Gln LysThr Gln Pro Asp Val Met Ser 3410 3415 3420 Gln Asn Ala Arg Lys Leu IleGln Lys Asn Leu Ala Thr Ser Ala Asp 3425 3430 3435 3440 Thr Pro Pro SerThr Val Pro Gly Thr Gly Lys Ser Val Ala Cys Ser 3445 3450 3455 Pro LysLys Ala Val Arg Asp Pro Lys Thr Gly Lys Ala Val Gln Glu 3460 3465 3470Arg Asn Ser Tyr Ala Val Ser Val Trp Lys Arg Val Lys Ala Lys Leu 34753480 3485 Glu Gly Arg Asp Val Asp Pro Asn Arg Arg Met Ser Val Ala GluGln 3490 3495 3500 Val Asp Tyr Val Ile Lys Glu Ala Thr Asn Leu Asp AsnLeu Ala Gln 3505 3510 3515 3520 Leu Tyr Glu Gly Trp Thr Ala Trp Val 35259 13110 DNA Homo sapiens CDS (328)...(11298) 9 ggggaagcag tggccgtgtgagcgtgagga gctgccgcca ccgcctgctc ctcgtcctcc 60 tcgtcctccg gggccccagcgtcgtgggcc gcgcacggcc ctggaagaga cgtcgcctcg 120 ccttcatccg cctctctcaccgcgccgctc cctcgtcctg ccctgcgggc tcaggcggaa 180 cccggaacgg ccgtcctcttcccccgccct ccgccgccgc ctcctcctcc tccttctcgg 240 cttcctcctc agccccgggccggagcgggg tgtcggcggc ggccggttcg ggcggcggcg 300 cttggccatg tcgtgtcggggaaggta atg agc cgc aga gcc ccg ggg tct cgg 354 Met Ser Arg Arg Ala ProGly Ser Arg 1 5 ctg agc agc ggc ggc acc aac tat tcg cgg agc tgg aat gactgg caa 402 Leu Ser Ser Gly Gly Thr Asn Tyr Ser Arg Ser Trp Asn Asp TrpGln 10 15 20 25 ccc aga act gat agt gca tca gct gac cca ggt aat tta aaatat tct 450 Pro Arg Thr Asp Ser Ala Ser Ala Asp Pro Gly Asn Leu Lys TyrSer 30 35 40 tca tcc aga gat aga ggt ggt tct tcc tct tac gga ctg caa ccttca 498 Ser Ser Arg Asp Arg Gly Gly Ser Ser Ser Tyr Gly Leu Gln Pro Ser45 50 55 aat tca gct gtg gtg tct cgg caa agg cac gat gat acc aga gtc cac546 Asn Ser Ala Val Val Ser Arg Gln Arg His Asp Asp Thr Arg Val His 6065 70 gct gac ata cag aat gac gaa aag ggt ggc tac agt gtc aat gga gga594 Ala Asp Ile Gln Asn Asp Glu Lys Gly Gly Tyr Ser Val Asn Gly Gly 7580 85 tct ggg gaa aat act tat ggt cgg aag tcg ttg ggg caa gag ctg agg642 Ser Gly Glu Asn Thr Tyr Gly Arg Lys Ser Leu Gly Gln Glu Leu Arg 9095 100 105 gtt aac aat gtg acc agc cct gag ttc acc agt gtt cag cat ggcagt 690 Val Asn Asn Val Thr Ser Pro Glu Phe Thr Ser Val Gln His Gly Ser110 115 120 cgt gct tta gcc acc aaa gac atg agg aaa tca cag gag aga tcgatg 738 Arg Ala Leu Ala Thr Lys Asp Met Arg Lys Ser Gln Glu Arg Ser Met125 130 135 tct tat tct gat gag tct cga ctg tcg aat ctt ctt cgg agg atcacc 786 Ser Tyr Ser Asp Glu Ser Arg Leu Ser Asn Leu Leu Arg Arg Ile Thr140 145 150 cgg gaa gac gac aga gac cga aga ttg gct act gta aag cag ttgaaa 834 Arg Glu Asp Asp Arg Asp Arg Arg Leu Ala Thr Val Lys Gln Leu Lys155 160 165 gaa ttt att cag caa cca gaa aat aag ctg gta cta gtt aaa caattg 882 Glu Phe Ile Gln Gln Pro Glu Asn Lys Leu Val Leu Val Lys Gln Leu170 175 180 185 gat aat atc ttg gct gct gta cat gac gtg ctt aat gaa agtagc aaa 930 Asp Asn Ile Leu Ala Ala Val His Asp Val Leu Asn Glu Ser SerLys 190 195 200 ttg ctt cag gag ttg aga cag gag gga gct tgc tgt ctt ggcctt ctt 978 Leu Leu Gln Glu Leu Arg Gln Glu Gly Ala Cys Cys Leu Gly LeuLeu 205 210 215 tgt gct tct ctg agc tat gag gct gag aag atc ttc aag tggatt ttt 1026 Cys Ala Ser Leu Ser Tyr Glu Ala Glu Lys Ile Phe Lys Trp IlePhe 220 225 230 agc aaa ttt agc tca tct gca aaa gat gaa gtt aaa ctc ctctac tta 1074 Ser Lys Phe Ser Ser Ser Ala Lys Asp Glu Val Lys Leu Leu TyrLeu 235 240 245 tgt gcc acc tac aaa gca cta gag act gta gga gaa aag aaagcc ttt 1122 Cys Ala Thr Tyr Lys Ala Leu Glu Thr Val Gly Glu Lys Lys AlaPhe 250 255 260 265 tca tct gta atg cag ctt gta atg acc agc ctg cag tctatt ctt gaa 1170 Ser Ser Val Met Gln Leu Val Met Thr Ser Leu Gln Ser IleLeu Glu 270 275 280 aat gtg gat aca cca gaa ttg ctt tgt aaa tgt gtt aagtgc att ctt 1218 Asn Val Asp Thr Pro Glu Leu Leu Cys Lys Cys Val Lys CysIle Leu 285 290 295 ttg gtg gct cga tgt tac cct cat att ttc agc act aatttt agg gat 1266 Leu Val Ala Arg Cys Tyr Pro His Ile Phe Ser Thr Asn PheArg Asp 300 305 310 aca gtt gat ata tta gtt gga tgg cat ata gat cat actcag aaa cct 1314 Thr Val Asp Ile Leu Val Gly Trp His Ile Asp His Thr GlnLys Pro 315 320 325 tcg ctc acg cag cag gta tct ggg tgg ttg cag agt ttggag cca ttt 1362 Ser Leu Thr Gln Gln Val Ser Gly Trp Leu Gln Ser Leu GluPro Phe 330 335 340 345 tgg gta gct gat ctt gca ttt tct act act ctt cttggt cag ttt ctg 1410 Trp Val Ala Asp Leu Ala Phe Ser Thr Thr Leu Leu GlyGln Phe Leu 350 355 360 gaa gac atg gaa gca tat gct gag gac ctc agc catgtg gcc tct ggg 1458 Glu Asp Met Glu Ala Tyr Ala Glu Asp Leu Ser His ValAla Ser Gly 365 370 375 gaa tca gtg gat gaa gat gtc cct cct cca tca gtgtca tta cca aag 1506 Glu Ser Val Asp Glu Asp Val Pro Pro Pro Ser Val SerLeu Pro Lys 380 385 390 ctg gct gca ctt ctc cgg gta ttt agt act gtg gtgagg agc att ggg 1554 Leu Ala Ala Leu Leu Arg Val Phe Ser Thr Val Val ArgSer Ile Gly 395 400 405 gaa cgc ttc agc cca att cgg ggt cct cca att actgag gca tat gta 1602 Glu Arg Phe Ser Pro Ile Arg Gly Pro Pro Ile Thr GluAla Tyr Val 410 415 420 425 aca gat gtt ctg tac aga gta atg aga tgt gtgacg gct gca aac cag 1650 Thr Asp Val Leu Tyr Arg Val Met Arg Cys Val ThrAla Ala Asn Gln 430 435 440 gtg ttt ttt tct gag gct gtg ttg aca gct gctaat gag tgt gtt ggt 1698 Val Phe Phe Ser Glu Ala Val Leu Thr Ala Ala AsnGlu Cys Val Gly 445 450 455 gtt ttg ctc ggc agc ttg gat cct agc atg actata cat tgt gac atg 1746 Val Leu Leu Gly Ser Leu Asp Pro Ser Met Thr IleHis Cys Asp Met 460 465 470 gtc att aca tat gga tta gac caa ctg gag aattgc cag act tgt ggt 1794 Val Ile Thr Tyr Gly Leu Asp Gln Leu Glu Asn CysGln Thr Cys Gly 475 480 485 acc gat tat atc atc tca gtc ttg aat tta ctcacg ctg att gtt gaa 1842 Thr Asp Tyr Ile Ile Ser Val Leu Asn Leu Leu ThrLeu Ile Val Glu 490 495 500 505 cag ata aat acg aaa ctg cca tca tca tttgta gaa aaa ctg ttt ata 1890 Gln Ile Asn Thr Lys Leu Pro Ser Ser Phe ValGlu Lys Leu Phe Ile 510 515 520 cca tca tct aaa cta cta ttc ttg cgt tatcat aaa gaa aaa gag gtt 1938 Pro Ser Ser Lys Leu Leu Phe Leu Arg Tyr HisLys Glu Lys Glu Val 525 530 535 gtt gct gta gcc cat gct gtt tat caa gcagtg ctc agc ttg aag aat 1986 Val Ala Val Ala His Ala Val Tyr Gln Ala ValLeu Ser Leu Lys Asn 540 545 550 att cct gtt ttg gag act gcc tat aag ttaata ttg gga gaa atg act 2034 Ile Pro Val Leu Glu Thr Ala Tyr Lys Leu IleLeu Gly Glu Met Thr 555 560 565 tgt gcc cta aac aac ctc cta cac agt ctacaa ctt cct gag gcc tgt 2082 Cys Ala Leu Asn Asn Leu Leu His Ser Leu GlnLeu Pro Glu Ala Cys 570 575 580 585 tct gaa ata aaa cat gag gct ttt aagaat cat gtg ttc aat gta gac 2130 Ser Glu Ile Lys His Glu Ala Phe Lys AsnHis Val Phe Asn Val Asp 590 595 600 aat gca aaa ttt gta gtt aaa ttt gacctc agt gcc ctg act aca att 2178 Asn Ala Lys Phe Val Val Lys Phe Asp LeuSer Ala Leu Thr Thr Ile 605 610 615 gga aat gcc aaa aac tca cta ata gggatg tgg gcg cta tct cca act 2226 Gly Asn Ala Lys Asn Ser Leu Ile Gly MetTrp Ala Leu Ser Pro Thr 620 625 630 gtc ttt gca ctt ctg agt aag aat ctgatg att gtg cac agt gac ctg 2274 Val Phe Ala Leu Leu Ser Lys Asn Leu MetIle Val His Ser Asp Leu 635 640 645 gct gtt cac ttc cct gcc att cag tatgct gtg ctc tac aca ttg tat 2322 Ala Val His Phe Pro Ala Ile Gln Tyr AlaVal Leu Tyr Thr Leu Tyr 650 655 660 665 tct cat tgt acc agg cat gat cacttt atc tct agt agc ctc agt tct 2370 Ser His Cys Thr Arg His Asp His PheIle Ser Ser Ser Leu Ser Ser 670 675 680 gcc tct cct tct ttg ttt gat ggagct gtg att agc act gta act acg 2418 Ala Ser Pro Ser Leu Phe Asp Gly AlaVal Ile Ser Thr Val Thr Thr 685 690 695 gct aca aag aaa cat ttc tca attata tta aat ctt ctg gga ata tta 2466 Ala Thr Lys Lys His Phe Ser Ile IleLeu Asn Leu Leu Gly Ile Leu 700 705 710 ctt aag aaa gat aac ctt aac caggac acg agg aaa ctg tta atg act 2514 Leu Lys Lys Asp Asn Leu Asn Gln AspThr Arg Lys Leu Leu Met Thr 715 720 725 tgg gct ttg gaa gca gct gtt ttaatg agg aag tct gaa aca tac gca 2562 Trp Ala Leu Glu Ala Ala Val Leu MetArg Lys Ser Glu Thr Tyr Ala 730 735 740 745 cct tta ttc tct ctt ccg tctttc cat aaa ttt tgc aaa ggc ctt tta 2610 Pro Leu Phe Ser Leu Pro Ser PheHis Lys Phe Cys Lys Gly Leu Leu 750 755 760 gcc aac act ctc gtt gaa gatgtg aat atc tgt ctg cag gca tgc agc 2658 Ala Asn Thr Leu Val Glu Asp ValAsn Ile Cys Leu Gln Ala Cys Ser 765 770 775 agt cta cat gct ctg tcc tcttcc ttg cca gat gat ctt tta cag aga 2706 Ser Leu His Ala Leu Ser Ser SerLeu Pro Asp Asp Leu Leu Gln Arg 780 785 790 tgt gtc gat gtt tgc cgt gttcaa cta gtg cac agt gga act cgt att 2754 Cys Val Asp Val Cys Arg Val GlnLeu Val His Ser Gly Thr Arg Ile 795 800 805 cga caa gca ttt gga aaa ctgttg aaa tca att cct tta gat gtt gtc 2802 Arg Gln Ala Phe Gly Lys Leu LeuLys Ser Ile Pro Leu Asp Val Val 810 815 820 825 cta agc aat aac aat cacaca gaa att caa gaa att tct tta gca tta 2850 Leu Ser Asn Asn Asn His ThrGlu Ile Gln Glu Ile Ser Leu Ala Leu 830 835 840 aga agt cac atg agt aaagca cca agt aat aca ttc cac ccc caa gat 2898 Arg Ser His Met Ser Lys AlaPro Ser Asn Thr Phe His Pro Gln Asp 845 850 855 ttc tct gat gtt att agtttt att ttg tat ggg aac tct cat aga aca 2946 Phe Ser Asp Val Ile Ser PheIle Leu Tyr Gly Asn Ser His Arg Thr 860 865 870 ggg aag gac aat tgg ttggaa aga ctg ttc tat agc tgc cag aga ctg 2994 Gly Lys Asp Asn Trp Leu GluArg Leu Phe Tyr Ser Cys Gln Arg Leu 875 880 885 gat aag cgt gac cag tcaaca att cca cgc aat ctc ctg aag aca gat 3042 Asp Lys Arg Asp Gln Ser ThrIle Pro Arg Asn Leu Leu Lys Thr Asp 890 895 900 905 gct gtc ctt tgg cagtgg gcc ata tgg gaa gct gca caa ttc act gtt 3090 Ala Val Leu Trp Gln TrpAla Ile Trp Glu Ala Ala Gln Phe Thr Val 910 915 920 ctt tct aag ctg agaacc cca ctg ggc aga gct caa gac acc ttc cag 3138 Leu Ser Lys Leu Arg ThrPro Leu Gly Arg Ala Gln Asp Thr Phe Gln 925 930 935 aca att gaa ggt atcatt cga agt ctc gca gct cac aca tta aac cct 3186 Thr Ile Glu Gly Ile IleArg Ser Leu Ala Ala His Thr Leu Asn Pro 940 945 950 gat cag gat gtt agtcag tgg aca act gca gac aat gat gaa ggc cat 3234 Asp Gln Asp Val Ser GlnTrp Thr Thr Ala Asp Asn Asp Glu Gly His 955 960 965 ggt aac aac caa cttaga ctt gtt ctt ctt ctg cag tat ctg gaa aat 3282 Gly Asn Asn Gln Leu ArgLeu Val Leu Leu Leu Gln Tyr Leu Glu Asn 970 975 980 985 ctg gag aaa ttaatg tat aat gca tac gag gga tgt gct aat gca tta 3330 Leu Glu Lys Leu MetTyr Asn Ala Tyr Glu Gly Cys Ala Asn Ala Leu 990 995 1000 act tca cct cccaag gtc att aga act ttt ttc tat acc aat cgc caa 3378 Thr Ser Pro Pro LysVal Ile Arg Thr Phe Phe Tyr Thr Asn Arg Gln 1005 1010 1015 act tgt caggac tgg cta acg cgg att cga ctc tcc atc atg agg gta 3426 Thr Cys Gln AspTrp Leu Thr Arg Ile Arg Leu Ser Ile Met Arg Val 1020 1025 1030 gga ttgttg gca ggc cag cct gca gtg aca gtg aga cat ggc ttt gac 3474 Gly Leu LeuAla Gly Gln Pro Ala Val Thr Val Arg His Gly Phe Asp 1035 1040 1045 ttgctt aca gag atg aaa aca acc agc cta tct cag ggg aat gaa ttg 3522 Leu LeuThr Glu Met Lys Thr Thr Ser Leu Ser Gln Gly Asn Glu Leu 1050 1055 10601065 gaa gta acc att atg atg gtg gta gaa gca tta tgt gaa ctt cat tgt3570 Glu Val Thr Ile Met Met Val Val Glu Ala Leu Cys Glu Leu His Cys1070 1075 1080 cct gaa gct ata cag gga att gct gtc tgg tca tca tct attgtt gga 3618 Pro Glu Ala Ile Gln Gly Ile Ala Val Trp Ser Ser Ser Ile ValGly 1085 1090 1095 aaa aat ctt ctg tgg att aac tca gtg gct caa cag gctgaa ggg agg 3666 Lys Asn Leu Leu Trp Ile Asn Ser Val Ala Gln Gln Ala GluGly Arg 1100 1105 1110 ttt gaa aag gcc tct gtg gag tac cag gaa cac ctgtgt gcc atg aca 3714 Phe Glu Lys Ala Ser Val Glu Tyr Gln Glu His Leu CysAla Met Thr 1115 1120 1125 ggt gtt gat tgc tgc atc tcc agc ttt gac aaatcg gtg ctc acc tta 3762 Gly Val Asp Cys Cys Ile Ser Ser Phe Asp Lys SerVal Leu Thr Leu 1130 1135 1140 1145 gcc aat gct ggg cgt aac agt gcc agcccg aaa cat tct ctg aat ggt 3810 Ala Asn Ala Gly Arg Asn Ser Ala Ser ProLys His Ser Leu Asn Gly 1150 1155 1160 gaa tcc aga aaa act gtg ctg tccaaa ccg act gac tct tcc cct gag 3858 Glu Ser Arg Lys Thr Val Leu Ser LysPro Thr Asp Ser Ser Pro Glu 1165 1170 1175 gtt ata aat tat tta gga aataaa gca tgt gag ttc tac atc tca att 3906 Val Ile Asn Tyr Leu Gly Asn LysAla Cys Glu Phe Tyr Ile Ser Ile 1180 1185 1190 gcc gat tgg gct gct gtgcag gaa tgg cag aac gct atc cat gac ttg 3954 Ala Asp Trp Ala Ala Val GlnGlu Trp Gln Asn Ala Ile His Asp Leu 1195 1200 1205 aaa aag agt acc agtagc act tcc ctc aac ctg aaa gct gac ttc aac 4002 Lys Lys Ser Thr Ser SerThr Ser Leu Asn Leu Lys Ala Asp Phe Asn 1210 1215 1220 1225 tat ata aaatca tta agc agc ttt gag tct gga aaa ttt gtt gaa tgt 4050 Tyr Ile Lys SerLeu Ser Ser Phe Glu Ser Gly Lys Phe Val Glu Cys 1230 1235 1240 acc gagcag tta gaa ttg tta cca gga gaa aat atc aat cta ctt gct 4098 Thr Glu GlnLeu Glu Leu Leu Pro Gly Glu Asn Ile Asn Leu Leu Ala 1245 1250 1255 ggagga tca aaa gaa aaa ata gac atg aaa aaa ctg ctt cct aac atg 4146 Gly GlySer Lys Glu Lys Ile Asp Met Lys Lys Leu Leu Pro Asn Met 1260 1265 1270tta agt ccg gat ccg agg gaa ctt cag aaa tcc att gaa gtt caa ttg 4194 LeuSer Pro Asp Pro Arg Glu Leu Gln Lys Ser Ile Glu Val Gln Leu 1275 12801285 tta aga agt tct gtt tgt ttg gca act gct tta aac ccg ata gaa caa4242 Leu Arg Ser Ser Val Cys Leu Ala Thr Ala Leu Asn Pro Ile Glu Gln1290 1295 1300 1305 gat cag aag tgg cag tct ata act gaa aat gtg gta aagtac ttg aag 4290 Asp Gln Lys Trp Gln Ser Ile Thr Glu Asn Val Val Lys TyrLeu Lys 1310 1315 1320 caa aca tcc cgc atc gct att gga cct ctg aga ctttct act tta aca 4338 Gln Thr Ser Arg Ile Ala Ile Gly Pro Leu Arg Leu SerThr Leu Thr 1325 1330 1335 gtt tca cag tct ttg cca gtt cta agt acc ttgcag ctg tat tgc tca 4386 Val Ser Gln Ser Leu Pro Val Leu Ser Thr Leu GlnLeu Tyr Cys Ser 1340 1345 1350 tct gct ttg gag aac aca gtt tct aac agactt tca aca gag gac tgt 4434 Ser Ala Leu Glu Asn Thr Val Ser Asn Arg LeuSer Thr Glu Asp Cys 1355 1360 1365 ctt att cca ctc ttc agt gaa gct ttacgt tca tgt aaa cag cat gac 4482 Leu Ile Pro Leu Phe Ser Glu Ala Leu ArgSer Cys Lys Gln His Asp 1370 1375 1380 1385 gtg agg cca tgg atg cag gcatta agg tat act atg tac cag aat cag 4530 Val Arg Pro Trp Met Gln Ala LeuArg Tyr Thr Met Tyr Gln Asn Gln 1390 1395 1400 ttg ttg gag aaa att aaagaa caa aca gtc cca att aga agc cat ctc 4578 Leu Leu Glu Lys Ile Lys GluGln Thr Val Pro Ile Arg Ser His Leu 1405 1410 1415 atg gaa tta ggt ctaaca gca gca aaa ttt gct aga aaa cga ggg aat 4626 Met Glu Leu Gly Leu ThrAla Ala Lys Phe Ala Arg Lys Arg Gly Asn 1420 1425 1430 gtg tcc ctt gcaaca aga ctg ctg gca cag tgc agt gaa gtt cag ctg 4674 Val Ser Leu Ala ThrArg Leu Leu Ala Gln Cys Ser Glu Val Gln Leu 1435 1440 1445 gga aag accacc act gca cag gat tta gtc caa cat ttt aaa aaa cta 4722 Gly Lys Thr ThrThr Ala Gln Asp Leu Val Gln His Phe Lys Lys Leu 1450 1455 1460 1465 tcaacc caa ggt caa gtg gat gaa aaa tgg ggg ccc gaa ctt gat att 4770 Ser ThrGln Gly Gln Val Asp Glu Lys Trp Gly Pro Glu Leu Asp Ile 1470 1475 1480gaa aaa acc aaa ttg ctt tat aca gca ggc cag tca aca cat gca atg 4818 GluLys Thr Lys Leu Leu Tyr Thr Ala Gly Gln Ser Thr His Ala Met 1485 14901495 gaa atg ttg agt tct tgt gcc ata tct ttc tgc aag tct gtg aaa gct4866 Glu Met Leu Ser Ser Cys Ala Ile Ser Phe Cys Lys Ser Val Lys Ala1500 1505 1510 gaa tat gca gtt gct aaa tca att ctg aca ctg gct aaa tggatc cag 4914 Glu Tyr Ala Val Ala Lys Ser Ile Leu Thr Leu Ala Lys Trp IleGln 1515 1520 1525 gca gaa tgg aaa gag att tca gga cag ctg aaa cag gtttac aga gct 4962 Ala Glu Trp Lys Glu Ile Ser Gly Gln Leu Lys Gln Val TyrArg Ala 1530 1535 1540 1545 cag cac caa cag aac ttc aca ggt ctt tct actttg tct aaa aac ata 5010 Gln His Gln Gln Asn Phe Thr Gly Leu Ser Thr LeuSer Lys Asn Ile 1550 1555 1560 ctc act cta ata gaa ctg cca tct gtt aatacg atg gaa gaa gag tat 5058 Leu Thr Leu Ile Glu Leu Pro Ser Val Asn ThrMet Glu Glu Glu Tyr 1565 1570 1575 cct cgg atc gag agt gaa tct aca gtgcat att gga gtt gga gaa cct 5106 Pro Arg Ile Glu Ser Glu Ser Thr Val HisIle Gly Val Gly Glu Pro 1580 1585 1590 gac ttc att ttg gga cag ttg tatcac ctg tct tca gta cag gca cct 5154 Asp Phe Ile Leu Gly Gln Leu Tyr HisLeu Ser Ser Val Gln Ala Pro 1595 1600 1605 gaa gta gcc aaa tct tgg gcagcg ttg gcc agc tgg gct tat agg tgg 5202 Glu Val Ala Lys Ser Trp Ala AlaLeu Ala Ser Trp Ala Tyr Arg Trp 1610 1615 1620 1625 ggc aga aag gtg gttgac aat gcc agt cag gga gaa ggt gtt cgt ctg 5250 Gly Arg Lys Val Val AspAsn Ala Ser Gln Gly Glu Gly Val Arg Leu 1630 1635 1640 ctg cct aga gaaaaa tct gaa gtt cag aat cta ctt cca gac act ata 5298 Leu Pro Arg Glu LysSer Glu Val Gln Asn Leu Leu Pro Asp Thr Ile 1645 1650 1655 act gag gaagag aaa gag aga ata tat ggt att ctt gga cag gct gtg 5346 Thr Glu Glu GluLys Glu Arg Ile Tyr Gly Ile Leu Gly Gln Ala Val 1660 1665 1670 tgt cggccg gcg ggg att cag gat gaa gat ata aca ctt cag ata act 5394 Cys Arg ProAla Gly Ile Gln Asp Glu Asp Ile Thr Leu Gln Ile Thr 1675 1680 1685 gagagt gaa gac aac gaa gaa gat gac atg gtt gat gtt atc tgg cgt 5442 Glu SerGlu Asp Asn Glu Glu Asp Asp Met Val Asp Val Ile Trp Arg 1690 1695 17001705 cag ttg ata tca agc tgc cca tgg ctt tca gaa ctt gat gaa agt gca5490 Gln Leu Ile Ser Ser Cys Pro Trp Leu Ser Glu Leu Asp Glu Ser Ala1710 1715 1720 act gaa gga gtt att aaa gtg tgg agg aaa gtt gta gat agaata ttc 5538 Thr Glu Gly Val Ile Lys Val Trp Arg Lys Val Val Asp Arg IlePhe 1725 1730 1735 agc ctg tac aaa ctc tct tgc agt gca tac ttt act ttcctt aaa ctc 5586 Ser Leu Tyr Lys Leu Ser Cys Ser Ala Tyr Phe Thr Phe LeuLys Leu 1740 1745 1750 aac gct ggt caa att cct tta gat gag gat gac cctagg ctg cat tta 5634 Asn Ala Gly Gln Ile Pro Leu Asp Glu Asp Asp Pro ArgLeu His Leu 1755 1760 1765 agt cac aga gtg gaa cag agc act gat gac atgatt gtg atg gcc aca 5682 Ser His Arg Val Glu Gln Ser Thr Asp Asp Met IleVal Met Ala Thr 1770 1775 1780 1785 ttg cgc ctg ctg cgg ttg ctc gtg aagcat gct ggt gag ctt cgg cag 5730 Leu Arg Leu Leu Arg Leu Leu Val Lys HisAla Gly Glu Leu Arg Gln 1790 1795 1800 tat ctg gag cac ggc ttg gag acaaca ccc act gca cca tgg agg gga 5778 Tyr Leu Glu His Gly Leu Glu Thr ThrPro Thr Ala Pro Trp Arg Gly 1805 1810 1815 att att ccg caa ctt ttc tcacgc tta aac cac cct gaa gtg tat gtg 5826 Ile Ile Pro Gln Leu Phe Ser ArgLeu Asn His Pro Glu Val Tyr Val 1820 1825 1830 cgc caa agt att tgt aacctt ctc tgc cgt gtg gct caa gat tcc cca 5874 Arg Gln Ser Ile Cys Asn LeuLeu Cys Arg Val Ala Gln Asp Ser Pro 1835 1840 1845 cat ctc ata ttg tatcct gca ata gtg ggt acc ata tcg ctt agt agt 5922 His Leu Ile Leu Tyr ProAla Ile Val Gly Thr Ile Ser Leu Ser Ser 1850 1855 1860 1865 gaa tcc caggct tca gga aat aaa ttt tcc act gca att cca act tta 5970 Glu Ser Gln AlaSer Gly Asn Lys Phe Ser Thr Ala Ile Pro Thr Leu 1870 1875 1880 ctt ggcaat att caa gga gaa gaa ttg ctg gtt tct gaa tgt gag gga 6018 Leu Gly AsnIle Gln Gly Glu Glu Leu Leu Val Ser Glu Cys Glu Gly 1885 1890 1895 ggaagt cct cct gca tct cag gat agc aat aag gat gaa cct aaa agt 6066 Gly SerPro Pro Ala Ser Gln Asp Ser Asn Lys Asp Glu Pro Lys Ser 1900 1905 1910gga tta aat gaa gac caa gcc atg atg cag gat tgt tac agc aaa att 6114 GlyLeu Asn Glu Asp Gln Ala Met Met Gln Asp Cys Tyr Ser Lys Ile 1915 19201925 gta gat aag ctg tcc tct gca aac ccc acc atg gta tta cag gtt cag6162 Val Asp Lys Leu Ser Ser Ala Asn Pro Thr Met Val Leu Gln Val Gln1930 1935 1940 1945 atg ctc gtg gct gaa ctg cgc agg gtc act gtg ctc tgggat gag ctc 6210 Met Leu Val Ala Glu Leu Arg Arg Val Thr Val Leu Trp AspGlu Leu 1950 1955 1960 tgg ctg gga gtt ttg ctg caa caa cac atg tat gtcctg aga cga att 6258 Trp Leu Gly Val Leu Leu Gln Gln His Met Tyr Val LeuArg Arg Ile 1965 1970 1975 cag cag ctt gaa gat gag gtg aag aga gtc cagaac aac aac acc tta 6306 Gln Gln Leu Glu Asp Glu Val Lys Arg Val Gln AsnAsn Asn Thr Leu 1980 1985 1990 cgc aaa gaa gag aaa att gca atc atg agggag agg cac aca gct ttg 6354 Arg Lys Glu Glu Lys Ile Ala Ile Met Arg GluArg His Thr Ala Leu 1995 2000 2005 atg aag ccc atc gta ttt gct ttg gagcat gtg agg agt atc aca gcg 6402 Met Lys Pro Ile Val Phe Ala Leu Glu HisVal Arg Ser Ile Thr Ala 2010 2015 2020 2025 gct cct gca gaa aca cct catgaa aaa tgg ttt cag gat aac tat ggt 6450 Ala Pro Ala Glu Thr Pro His GluLys Trp Phe Gln Asp Asn Tyr Gly 2030 2035 2040 gat gcc att gaa aat gcccta gaa aaa ctg aag act cca ttg aac cct 6498 Asp Ala Ile Glu Asn Ala LeuGlu Lys Leu Lys Thr Pro Leu Asn Pro 2045 2050 2055 gca aag cct ggg agcagc tgg att cca ttt aaa gag ata atg cta agt 6546 Ala Lys Pro Gly Ser SerTrp Ile Pro Phe Lys Glu Ile Met Leu Ser 2060 2065 2070 ttg caa cag agagca cag aaa cgt gca agt tac atc ttg cgt ctt gaa 6594 Leu Gln Gln Arg AlaGln Lys Arg Ala Ser Tyr Ile Leu Arg Leu Glu 2075 2080 2085 gaa atc agtcca tgg ttg gct gcc atg act aac act gaa att gct ctt 6642 Glu Ile Ser ProTrp Leu Ala Ala Met Thr Asn Thr Glu Ile Ala Leu 2090 2095 2100 2105 cctggg gaa gtc tca gcc aga gac act gtc aca atc cat agt gtg ggc 6690 Pro GlyGlu Val Ser Ala Arg Asp Thr Val Thr Ile His Ser Val Gly 2110 2115 2120gga acc atc aca atc tta ccg act aaa acc aag cca aag aaa ctt ctc 6738 GlyThr Ile Thr Ile Leu Pro Thr Lys Thr Lys Pro Lys Lys Leu Leu 2125 21302135 ttt ctt gga tca gat ggg aag agc tat cct tat ctt ttc aaa gga ctg6786 Phe Leu Gly Ser Asp Gly Lys Ser Tyr Pro Tyr Leu Phe Lys Gly Leu2140 2145 2150 gag gat tta cat ctg gat gag aga ata atg cag ttc cta tctatt gtg 6834 Glu Asp Leu His Leu Asp Glu Arg Ile Met Gln Phe Leu Ser IleVal 2155 2160 2165 aat acc atg ttt gct aca att aat cgc caa gaa aca ccccgg ttc cat 6882 Asn Thr Met Phe Ala Thr Ile Asn Arg Gln Glu Thr Pro ArgPhe His 2170 2175 2180 2185 gct cga cac tat tct gta aca cca cta gga acaaga tca gga cta atc 6930 Ala Arg His Tyr Ser Val Thr Pro Leu Gly Thr ArgSer Gly Leu Ile 2190 2195 2200 cag tgg gta gat gga gcc aca ccc tta tttggt ctt tac aaa cga tgg 6978 Gln Trp Val Asp Gly Ala Thr Pro Leu Phe GlyLeu Tyr Lys Arg Trp 2205 2210 2215 caa caa cgg gaa gct gcc tta caa gcacaa aag gcc caa gat tcc tac 7026 Gln Gln Arg Glu Ala Ala Leu Gln Ala GlnLys Ala Gln Asp Ser Tyr 2220 2225 2230 caa act cct cag aat cct gga attgta ccc cgt cct agt gaa ctt tat 7074 Gln Thr Pro Gln Asn Pro Gly Ile ValPro Arg Pro Ser Glu Leu Tyr 2235 2240 2245 tac agt aaa att ggc cct gctttg aaa aca gtt ggg ctt agc ctg gat 7122 Tyr Ser Lys Ile Gly Pro Ala LeuLys Thr Val Gly Leu Ser Leu Asp 2250 2255 2260 2265 gtg tcc cgt cgg gattgg cct ctt cat gta atg aag gca gta ttg gaa 7170 Val Ser Arg Arg Asp TrpPro Leu His Val Met Lys Ala Val Leu Glu 2270 2275 2280 gag tta atg gaggcc aca ccc ccg aat ctc ctt gcc aaa gag ctc tgg 7218 Glu Leu Met Glu AlaThr Pro Pro Asn Leu Leu Ala Lys Glu Leu Trp 2285 2290 2295 tca tct tgcaca aca cct gat gaa tgg tgg aga gtt acg cag tct tat 7266 Ser Ser Cys ThrThr Pro Asp Glu Trp Trp Arg Val Thr Gln Ser Tyr 2300 2305 2310 gca agatct act gca gtc atg tct atg gtt gga tac ata att ggc ctt 7314 Ala Arg SerThr Ala Val Met Ser Met Val Gly Tyr Ile Ile Gly Leu 2315 2320 2325 ggagac aga cat ctg gat aat gtt ctt ata gat atg acg act gga gaa 7362 Gly AspArg His Leu Asp Asn Val Leu Ile Asp Met Thr Thr Gly Glu 2330 2335 23402345 gtt gtt cac ata gat tac aat gtt tgc ttt gaa aaa ggt aaa agc ctt7410 Val Val His Ile Asp Tyr Asn Val Cys Phe Glu Lys Gly Lys Ser Leu2350 2355 2360 aga gtt cct gag aaa gta cct ttt cga atg aca caa aac attgaa aca 7458 Arg Val Pro Glu Lys Val Pro Phe Arg Met Thr Gln Asn Ile GluThr 2365 2370 2375 gca ctg ggt gta act gga gta gaa ggt gta ttt agg ctttca tgt gag 7506 Ala Leu Gly Val Thr Gly Val Glu Gly Val Phe Arg Leu SerCys Glu 2380 2385 2390 cag gtt tta cac att atg cgg cgt ggc aga gag accctg ctg acg ctg 7554 Gln Val Leu His Ile Met Arg Arg Gly Arg Glu Thr LeuLeu Thr Leu 2395 2400 2405 ctg gag gcc ttt gtg tac gac cct ctg gtg gactgg aca gca gga ggc 7602 Leu Glu Ala Phe Val Tyr Asp Pro Leu Val Asp TrpThr Ala Gly Gly 2410 2415 2420 2425 gag gct ggg ttt gct ggt gct gtc tatggt gga ggt ggc cag cag gcc 7650 Glu Ala Gly Phe Ala Gly Ala Val Tyr GlyGly Gly Gly Gln Gln Ala 2430 2435 2440 gag agc aag cag agc aag aga gagatg gag cga gag atc acc cgc agc 7698 Glu Ser Lys Gln Ser Lys Arg Glu MetGlu Arg Glu Ile Thr Arg Ser 2445 2450 2455 ctg ttt tct tct aga gta gctgag att aag gtg aac tgg ttt aag aat 7746 Leu Phe Ser Ser Arg Val Ala GluIle Lys Val Asn Trp Phe Lys Asn 2460 2465 2470 aga gat gag atg ctg gttgtg ctt ccc aag ttg gac ggt agc tta gat 7794 Arg Asp Glu Met Leu Val ValLeu Pro Lys Leu Asp Gly Ser Leu Asp 2475 2480 2485 gaa tac cta agc ttgcaa gag caa ctg aca gat gtg gaa aaa ctg cag 7842 Glu Tyr Leu Ser Leu GlnGlu Gln Leu Thr Asp Val Glu Lys Leu Gln 2490 2495 2500 2505 ggc aaa ctactg gag gaa ata gag ttt cta gaa gga gct gaa ggg gtg 7890 Gly Lys Leu LeuGlu Glu Ile Glu Phe Leu Glu Gly Ala Glu Gly Val 2510 2515 2520 gat catcct tct cat act ctg caa cac agg tat tct gag cac acc caa 7938 Asp His ProSer His Thr Leu Gln His Arg Tyr Ser Glu His Thr Gln 2525 2530 2535 ctacag act cag caa aga gct gtt cag gaa gca atc cag gtg aag ctg 7986 Leu GlnThr Gln Gln Arg Ala Val Gln Glu Ala Ile Gln Val Lys Leu 2540 2545 2550aat gaa ttt gaa caa tgg ata aca cat tat cag gct gca ttc aat aat 8034 AsnGlu Phe Glu Gln Trp Ile Thr His Tyr Gln Ala Ala Phe Asn Asn 2555 25602565 tta gaa gca aca cag ctt gca agc ttg ctt caa gag ata agc aca caa8082 Leu Glu Ala Thr Gln Leu Ala Ser Leu Leu Gln Glu Ile Ser Thr Gln2570 2575 2580 2585 atg gac ctt ggt cct cca agt tac gtg cca gca aca gccttt ctg cag 8130 Met Asp Leu Gly Pro Pro Ser Tyr Val Pro Ala Thr Ala PheLeu Gln 2590 2595 2600 aat gct ggt cag gcc cac ttg att agc cag tgc gagcag ctg gag ggg 8178 Asn Ala Gly Gln Ala His Leu Ile Ser Gln Cys Glu GlnLeu Glu Gly 2605 2610 2615 gag gtt ggt gct ctc ctg cag cag agg cgc tccgtg ctc cgt ggc tgt 8226 Glu Val Gly Ala Leu Leu Gln Gln Arg Arg Ser ValLeu Arg Gly Cys 2620 2625 2630 ctg gag caa ctg cat cac tat gca acc gtggcc ctg cag tat ccg aag 8274 Leu Glu Gln Leu His His Tyr Ala Thr Val AlaLeu Gln Tyr Pro Lys 2635 2640 2645 gcc ata ttt cag aaa cat cga att gaacag tgg aag acc tgg atg gaa 8322 Ala Ile Phe Gln Lys His Arg Ile Glu GlnTrp Lys Thr Trp Met Glu 2650 2655 2660 2665 gag ctc atc tgt aac acc acagta gag cgt tgt caa gag ctc tat agg 8370 Glu Leu Ile Cys Asn Thr Thr ValGlu Arg Cys Gln Glu Leu Tyr Arg 2670 2675 2680 aaa tat gaa atg caa tatgct ccc cag cca ccc cca aca gtg tgt cag 8418 Lys Tyr Glu Met Gln Tyr AlaPro Gln Pro Pro Pro Thr Val Cys Gln 2685 2690 2695 ttc atc act gcc actgaa atg acc ctg cag cga tac gca gca gac atc 8466 Phe Ile Thr Ala Thr GluMet Thr Leu Gln Arg Tyr Ala Ala Asp Ile 2700 2705 2710 aac agc aga cttatt aga caa gtg gaa cgc ttg aaa cag gaa gct gtc 8514 Asn Ser Arg Leu IleArg Gln Val Glu Arg Leu Lys Gln Glu Ala Val 2715 2720 2725 act gtg ccagtt tgt gaa gat cag ttg aaa gaa att gaa cgt tgc att 8562 Thr Val Pro ValCys Glu Asp Gln Leu Lys Glu Ile Glu Arg Cys Ile 2730 2735 2740 2745 aaagtt ttc ctt cat gag aat gga gaa gaa gga tct ttg agt cta gca 8610 Lys ValPhe Leu His Glu Asn Gly Glu Glu Gly Ser Leu Ser Leu Ala 2750 2755 2760agt gtt att att tct gcc ctt tgt acc ctt aca agg cgt aac ctg atg 8658 SerVal Ile Ile Ser Ala Leu Cys Thr Leu Thr Arg Arg Asn Leu Met 2765 27702775 atg gaa ggt gca gcg tca agt gct gga gaa cag ctg gtt gat ctg act8706 Met Glu Gly Ala Ala Ser Ser Ala Gly Glu Gln Leu Val Asp Leu Thr2780 2785 2790 tct cgg gat gga gcc tgg ttc ttg gag gaa ctc tgc agt atgagc gga 8754 Ser Arg Asp Gly Ala Trp Phe Leu Glu Glu Leu Cys Ser Met SerGly 2795 2800 2805 aac gtc acc tgc ttg gtt cag tta ctg aag cag tgc cacctg gtg cca 8802 Asn Val Thr Cys Leu Val Gln Leu Leu Lys Gln Cys His LeuVal Pro 2810 2815 2820 2825 cag gac tta gat atc ccg aac ccc atg gaa gcgtct gag aca gtt cac 8850 Gln Asp Leu Asp Ile Pro Asn Pro Met Glu Ala SerGlu Thr Val His 2830 2835 2840 tta gcc aat gga gtg tat acc tca ctt caggaa ttg aat tcg aat ttc 8898 Leu Ala Asn Gly Val Tyr Thr Ser Leu Gln GluLeu Asn Ser Asn Phe 2845 2850 2855 cgg caa atc ata ttt cca gaa gca cttcga tgt tta atg aaa ggg gaa 8946 Arg Gln Ile Ile Phe Pro Glu Ala Leu ArgCys Leu Met Lys Gly Glu 2860 2865 2870 tac acg tta gaa agt atg ctg catgaa ctg gac ggt ctt att gag cag 8994 Tyr Thr Leu Glu Ser Met Leu His GluLeu Asp Gly Leu Ile Glu Gln 2875 2880 2885 acc acc gat ggc gtt ccc ctgcag act cta gtg gaa tct ctt cag gcc 9042 Thr Thr Asp Gly Val Pro Leu GlnThr Leu Val Glu Ser Leu Gln Ala 2890 2895 2900 2905 tac tta aga aac gcagct atg gga ctg gaa gaa gaa aca cat gct cat 9090 Tyr Leu Arg Asn Ala AlaMet Gly Leu Glu Glu Glu Thr His Ala His 2910 2915 2920 tac atc gat gttgcc aga cta cta cat gct cag tac ggt gaa tta atc 9138 Tyr Ile Asp Val AlaArg Leu Leu His Ala Gln Tyr Gly Glu Leu Ile 2925 2930 2935 caa ccg agaaat ggt tca gtt gat gaa aca ccc aaa atg tca gct ggc 9186 Gln Pro Arg AsnGly Ser Val Asp Glu Thr Pro Lys Met Ser Ala Gly 2940 2945 2950 cag atgctt ttg gta gca ttc gat ggc atg ttt gct caa gtt gaa act 9234 Gln Met LeuLeu Val Ala Phe Asp Gly Met Phe Ala Gln Val Glu Thr 2955 2960 2965 gctttc agc tta tta gtt gaa aag ttg aac aag atg gaa att ccc ata 9282 Ala PheSer Leu Leu Val Glu Lys Leu Asn Lys Met Glu Ile Pro Ile 2970 2975 29802985 gct tgg cga aag att gac atc ata agg gaa gcc agg agt act caa gtt9330 Ala Trp Arg Lys Ile Asp Ile Ile Arg Glu Ala Arg Ser Thr Gln Val2990 2995 3000 aat ttt ttt gat gat gat aat cac cgg cag gtg cta gaa gagatt ttc 9378 Asn Phe Phe Asp Asp Asp Asn His Arg Gln Val Leu Glu Glu IlePhe 3005 3010 3015 ttt cta aaa aga cta cag act att aag gag ttc ttc aggctc tgt ggt 9426 Phe Leu Lys Arg Leu Gln Thr Ile Lys Glu Phe Phe Arg LeuCys Gly 3020 3025 3030 acc ttt tct aaa aca ttg tca gga tca agt tca cttgaa gat cag aat 9474 Thr Phe Ser Lys Thr Leu Ser Gly Ser Ser Ser Leu GluAsp Gln Asn 3035 3040 3045 act gtg aat ggg cct gta cag att gtc aat gtgaaa acc ctt ttt aga 9522 Thr Val Asn Gly Pro Val Gln Ile Val Asn Val LysThr Leu Phe Arg 3050 3055 3060 3065 aac tct tgt ttc agt gaa gac caa atggcc aaa cct atc aag gca ttc 9570 Asn Ser Cys Phe Ser Glu Asp Gln Met AlaLys Pro Ile Lys Ala Phe 3070 3075 3080 aca gct gac ttt gtg agg cag ctcttg ata ggg cta ccc aac caa gcc 9618 Thr Ala Asp Phe Val Arg Gln Leu LeuIle Gly Leu Pro Asn Gln Ala 3085 3090 3095 ctc gga ctc aca ctg tgc agtttt atc agt gct ctg ggt gta gac atc 9666 Leu Gly Leu Thr Leu Cys Ser PheIle Ser Ala Leu Gly Val Asp Ile 3100 3105 3110 att gct caa gta gag gcaaag gac ttt ggt gcc gaa agc aaa gtt tct 9714 Ile Ala Gln Val Glu Ala LysAsp Phe Gly Ala Glu Ser Lys Val Ser 3115 3120 3125 gtt gat gat ctc tgtaag aaa gcg gtg gaa cat aac atc cag ata ggg 9762 Val Asp Asp Leu Cys LysLys Ala Val Glu His Asn Ile Gln Ile Gly 3130 3135 3140 3145 aag ttc tctcag ctg gtt atg aac agg gca act gtg tta gca agt tct 9810 Lys Phe Ser GlnLeu Val Met Asn Arg Ala Thr Val Leu Ala Ser Ser 3150 3155 3160 tac gacact gcc tgg aag aag cat gac ttg gtg cga agg cta gaa acc 9858 Tyr Asp ThrAla Trp Lys Lys His Asp Leu Val Arg Arg Leu Glu Thr 3165 3170 3175 agtatt tct tct tgt aag aca agc ctg cag cgg gtt cag ctg cat att 9906 Ser IleSer Ser Cys Lys Thr Ser Leu Gln Arg Val Gln Leu His Ile 3180 3185 3190gcc atg ttt cag tgg caa cat gaa gat cta ctt atc aat aga cca caa 9954 AlaMet Phe Gln Trp Gln His Glu Asp Leu Leu Ile Asn Arg Pro Gln 3195 32003205 gcc atg tca gtc aca cct ccc cca cgg tct gct atc cta acc agc atg10002 Ala Met Ser Val Thr Pro Pro Pro Arg Ser Ala Ile Leu Thr Ser Met3210 3215 3220 3225 aaa aag aag ctg cat acc ctg agc cag att gaa act tctatt gcg aca 10050 Lys Lys Lys Leu His Thr Leu Ser Gln Ile Glu Thr SerIle Ala Thr 3230 3235 3240 gtt cag gag aag cta gct gca ctt gaa tca agtatt gaa cag cga ctc 10098 Val Gln Glu Lys Leu Ala Ala Leu Glu Ser SerIle Glu Gln Arg Leu 3245 3250 3255 aag tgg gca ggt ggt gcc aac cct gcattg gcc cct gta cta caa gat 10146 Lys Trp Ala Gly Gly Ala Asn Pro AlaLeu Ala Pro Val Leu Gln Asp 3260 3265 3270 ttt gaa gca acg ata gct gaaaga aga aat ctt gtc ctt aaa gag agc 10194 Phe Glu Ala Thr Ile Ala GluArg Arg Asn Leu Val Leu Lys Glu Ser 3275 3280 3285 caa aga gca agt caggtc aca ttt ctc tgc agc aat atc att cat ttt 10242 Gln Arg Ala Ser GlnVal Thr Phe Leu Cys Ser Asn Ile Ile His Phe 3290 3295 3300 3305 gaa agttta cga aca aga act gca gaa gcc tta aac ctg gat gcg gcg 10290 Glu SerLeu Arg Thr Arg Thr Ala Glu Ala Leu Asn Leu Asp Ala Ala 3310 3315 3320tta ttt gaa cta atc aag cga tgt cag cag atg tgt tcg ttt gca tca 10338Leu Phe Glu Leu Ile Lys Arg Cys Gln Gln Met Cys Ser Phe Ala Ser 33253330 3335 cag ttt aac agt tca gtg tct gag tta gag ctt cgt tta tta cagaga 10386 Gln Phe Asn Ser Ser Val Ser Glu Leu Glu Leu Arg Leu Leu GlnArg 3340 3345 3350 gtg gac act ggt ctt gaa cat cct att ggc agc tct gaatgg ctt ttg 10434 Val Asp Thr Gly Leu Glu His Pro Ile Gly Ser Ser GluTrp Leu Leu 3355 3360 3365 tca gca cac aaa cag ttg acc cag gat atg tctact cag agg gca att 10482 Ser Ala His Lys Gln Leu Thr Gln Asp Met SerThr Gln Arg Ala Ile 3370 3375 3380 3385 cag aca gag aaa gag cag cag atagaa acg gtc tgt gaa aca att cag 10530 Gln Thr Glu Lys Glu Gln Gln IleGlu Thr Val Cys Glu Thr Ile Gln 3390 3395 3400 aat ctg gtt gat aat ataaag act gtg ctc act ggt cat aac cga cag 10578 Asn Leu Val Asp Asn IleLys Thr Val Leu Thr Gly His Asn Arg Gln 3405 3410 3415 ctt gga gat gtcaaa cat ctc ttg aaa gct atg gct aag gat gaa gaa 10626 Leu Gly Asp ValLys His Leu Leu Lys Ala Met Ala Lys Asp Glu Glu 3420 3425 3430 gct gctctg gca gat ggt gaa gat gtt ccc tat gag aac agt gtt agg 10674 Ala AlaLeu Ala Asp Gly Glu Asp Val Pro Tyr Glu Asn Ser Val Arg 3435 3440 3445cag ttt ttg ggt gaa tat aaa tca tgg caa gac aac att caa aca gtt 10722Gln Phe Leu Gly Glu Tyr Lys Ser Trp Gln Asp Asn Ile Gln Thr Val 34503455 3460 3465 cta ttt aca tta gtc cag gct atg ggt cag gtt cga agt caagaa cac 10770 Leu Phe Thr Leu Val Gln Ala Met Gly Gln Val Arg Ser GlnGlu His 3470 3475 3480 gtt gaa atg ctc cag gaa atc act ccc acc ttg aaagaa ctg aaa aca 10818 Val Glu Met Leu Gln Glu Ile Thr Pro Thr Leu LysGlu Leu Lys Thr 3485 3490 3495 caa agt cag agt atc tat aat aat tta gtgagt ttt gca tca ccc tta 10866 Gln Ser Gln Ser Ile Tyr Asn Asn Leu ValSer Phe Ala Ser Pro Leu 3500 3505 3510 gtc acc gat gca aca aat gaa tgttcg agt cca acg tca tct gct act 10914 Val Thr Asp Ala Thr Asn Glu CysSer Ser Pro Thr Ser Ser Ala Thr 3515 3520 3525 tat cag cca tcc ttc gctgca gca gtc cgg agt aac act ggc cag aag 10962 Tyr Gln Pro Ser Phe AlaAla Ala Val Arg Ser Asn Thr Gly Gln Lys 3530 3535 3540 3545 act cag cctgat gtc atg tca cag aat gct aga aag ctg atc cag aaa 11010 Thr Gln ProAsp Val Met Ser Gln Asn Ala Arg Lys Leu Ile Gln Lys 3550 3555 3560 aatctt gct aca tca gct gat act cca cca agc acc gtt cca gga act 11058 AsnLeu Ala Thr Ser Ala Asp Thr Pro Pro Ser Thr Val Pro Gly Thr 3565 35703575 ggc aag agt gtt gct tgt agt cct aaa aag gca gtc aga gac cct aaa11106 Gly Lys Ser Val Ala Cys Ser Pro Lys Lys Ala Val Arg Asp Pro Lys3580 3585 3590 act ggg aaa gcg gtg caa gag aga aac tcc tat gca gtg agtgtg tgg 11154 Thr Gly Lys Ala Val Gln Glu Arg Asn Ser Tyr Ala Val SerVal Trp 3595 3600 3605 aag aga gtg aaa gcc aag tta gag ggc cga gat gttgat ccg aat agg 11202 Lys Arg Val Lys Ala Lys Leu Glu Gly Arg Asp ValAsp Pro Asn Arg 3610 3615 3620 3625 agg atg tca gtt gct gaa cag gtt gactat gtc att aag gaa gca act 11250 Arg Met Ser Val Ala Glu Gln Val AspTyr Val Ile Lys Glu Ala Thr 3630 3635 3640 aat cta gat aac ttg gct cagctg tat gaa ggt tgg aca gcc tgg gtg 11298 Asn Leu Asp Asn Leu Ala GlnLeu Tyr Glu Gly Trp Thr Ala Trp Val 3645 3650 3655 tgaatggcaa gacagtagatgagtctggtt aagcgaggtc agacatccac cagaatcaac 11358 tcagcctcag gcatccaaagccacaccaca gtcggtggtg atgcaactgg gggcttactc 11418 tgaggaaacc taggaaatctcggtgcacta ggaagtgaat cccgcaggac agctgcactc 11478 agggatacgc ccaacaccatggcctgcaac cccagggtca agggtgaagg aaagcaaagc 11538 tcaccgcctg aacacggagattgtctttct gccacagaac agcagcagac gtgtcgggag 11598 gttagctgcg gaaagaaatcgggatgccgc ggagcacaga gtgatttgga actccattcc 11658 acctgaccct gtgtgtacaatccaggaaaa aaacaaaccc cactcagaaa cagagaaaac 11718 tggggtcgcg aagaaatcacagccaaggaa gatttgatgc attcagattc tcgtgtaaca 11778 cttgttgctt ggcaacagtactggttgggt tgaccagtaa gtagaaaaag gctaaaggct 11838 atgcgatatg aatttcagaaatggactgaa aatggagagc tatgtaacag atacactaca 11898 gtagaagaac ttacttctgaaatgaaggga aaaaaaccac cccatcgttc cctactcctc 11958 cccaccactt acccgttccccctttaccta atctagtaga ttagccatct ttcaaattca 12018 cttttatttc agtccttatatttcatatac ttccgtctcg atgctgttaa caacttctga 12078 taacatggaa aattcaaggattgtttaaag gtctgatgat cacacacaaa atgtaattcc 12138 ggttatttaa gtcatttctgtgattctatc atgtacagtt tccagaattg tcactgtgca 12198 ttcaaaagta atgaatctaacagacatttg atttaatgta cactcccttt tgcttatagt 12258 gtgcattttt tttggaggtcattcaaattt tccctcttct gtgatagctg tagtttcttt 12318 catagaaagt agctaatccagtgtaatctt ttaccttttt aaaaaccaag atagagtatc 12378 tattagagtt ttacattgttgatgatagat taacaataaa gtgatgttct ggtggaggta 12438 gactgaaatt tttttaattcatgtttttca tttgatactt ttaatttaca cttagtaaat 12498 taaaagttgt ttaatttacttggcatttta ggacatgtac atgaaacagt gaaaatgaga 12558 tccaccaaca tcttttattaagttcagtta ttagtctgtg aagtgcttta ctttttgcac 12618 aattttaata gcttgctattcagtaataca ttatagtgaa ttcatgatca aggtttcctt 12678 aaatttagca ttgcatttcagtactgactg tgtaagctaa attgctgatc caaaataaaa 12738 acccagacta gaatagggttcttaaaatca agtatcaata caaaatagaa cacaattaaa 12798 atcttaattg ttggctgggcacagtggctc acgcctgtaa tcccagcact ttgggaggcc 12858 gaggcgggcg gatcatgaggttaggagagc gagaccatcc tggctaacac ggtgaaaccc 12918 cgtctttact aaaatacaaaaaaaattagc cgggtgtggt ggcgggcgcc tgtagtccca 12978 gctactcggg aggctgaggcaggagaatgg cgtgaaccca ggaggcggag cttgcagtga 13038 gccgagattg tgccactgcactccagcctg ggcaacagag ctagactctg tgtcaaaaat 13098 aaatgactag at 13110 103657 PRT Homo sapiens 10 Met Ser Arg Arg Ala Pro Gly Ser Arg Leu Ser SerGly Gly Thr Asn 1 5 10 15 Tyr Ser Arg Ser Trp Asn Asp Trp Gln Pro ArgThr Asp Ser Ala Ser 20 25 30 Ala Asp Pro Gly Asn Leu Lys Tyr Ser Ser SerArg Asp Arg Gly Gly 35 40 45 Ser Ser Ser Tyr Gly Leu Gln Pro Ser Asn SerAla Val Val Ser Arg 50 55 60 Gln Arg His Asp Asp Thr Arg Val His Ala AspIle Gln Asn Asp Glu 65 70 75 80 Lys Gly Gly Tyr Ser Val Asn Gly Gly SerGly Glu Asn Thr Tyr Gly 85 90 95 Arg Lys Ser Leu Gly Gln Glu Leu Arg ValAsn Asn Val Thr Ser Pro 100 105 110 Glu Phe Thr Ser Val Gln His Gly SerArg Ala Leu Ala Thr Lys Asp 115 120 125 Met Arg Lys Ser Gln Glu Arg SerMet Ser Tyr Ser Asp Glu Ser Arg 130 135 140 Leu Ser Asn Leu Leu Arg ArgIle Thr Arg Glu Asp Asp Arg Asp Arg 145 150 155 160 Arg Leu Ala Thr ValLys Gln Leu Lys Glu Phe Ile Gln Gln Pro Glu 165 170 175 Asn Lys Leu ValLeu Val Lys Gln Leu Asp Asn Ile Leu Ala Ala Val 180 185 190 His Asp ValLeu Asn Glu Ser Ser Lys Leu Leu Gln Glu Leu Arg Gln 195 200 205 Glu GlyAla Cys Cys Leu Gly Leu Leu Cys Ala Ser Leu Ser Tyr Glu 210 215 220 AlaGlu Lys Ile Phe Lys Trp Ile Phe Ser Lys Phe Ser Ser Ser Ala 225 230 235240 Lys Asp Glu Val Lys Leu Leu Tyr Leu Cys Ala Thr Tyr Lys Ala Leu 245250 255 Glu Thr Val Gly Glu Lys Lys Ala Phe Ser Ser Val Met Gln Leu Val260 265 270 Met Thr Ser Leu Gln Ser Ile Leu Glu Asn Val Asp Thr Pro GluLeu 275 280 285 Leu Cys Lys Cys Val Lys Cys Ile Leu Leu Val Ala Arg CysTyr Pro 290 295 300 His Ile Phe Ser Thr Asn Phe Arg Asp Thr Val Asp IleLeu Val Gly 305 310 315 320 Trp His Ile Asp His Thr Gln Lys Pro Ser LeuThr Gln Gln Val Ser 325 330 335 Gly Trp Leu Gln Ser Leu Glu Pro Phe TrpVal Ala Asp Leu Ala Phe 340 345 350 Ser Thr Thr Leu Leu Gly Gln Phe LeuGlu Asp Met Glu Ala Tyr Ala 355 360 365 Glu Asp Leu Ser His Val Ala SerGly Glu Ser Val Asp Glu Asp Val 370 375 380 Pro Pro Pro Ser Val Ser LeuPro Lys Leu Ala Ala Leu Leu Arg Val 385 390 395 400 Phe Ser Thr Val ValArg Ser Ile Gly Glu Arg Phe Ser Pro Ile Arg 405 410 415 Gly Pro Pro IleThr Glu Ala Tyr Val Thr Asp Val Leu Tyr Arg Val 420 425 430 Met Arg CysVal Thr Ala Ala Asn Gln Val Phe Phe Ser Glu Ala Val 435 440 445 Leu ThrAla Ala Asn Glu Cys Val Gly Val Leu Leu Gly Ser Leu Asp 450 455 460 ProSer Met Thr Ile His Cys Asp Met Val Ile Thr Tyr Gly Leu Asp 465 470 475480 Gln Leu Glu Asn Cys Gln Thr Cys Gly Thr Asp Tyr Ile Ile Ser Val 485490 495 Leu Asn Leu Leu Thr Leu Ile Val Glu Gln Ile Asn Thr Lys Leu Pro500 505 510 Ser Ser Phe Val Glu Lys Leu Phe Ile Pro Ser Ser Lys Leu LeuPhe 515 520 525 Leu Arg Tyr His Lys Glu Lys Glu Val Val Ala Val Ala HisAla Val 530 535 540 Tyr Gln Ala Val Leu Ser Leu Lys Asn Ile Pro Val LeuGlu Thr Ala 545 550 555 560 Tyr Lys Leu Ile Leu Gly Glu Met Thr Cys AlaLeu Asn Asn Leu Leu 565 570 575 His Ser Leu Gln Leu Pro Glu Ala Cys SerGlu Ile Lys His Glu Ala 580 585 590 Phe Lys Asn His Val Phe Asn Val AspAsn Ala Lys Phe Val Val Lys 595 600 605 Phe Asp Leu Ser Ala Leu Thr ThrIle Gly Asn Ala Lys Asn Ser Leu 610 615 620 Ile Gly Met Trp Ala Leu SerPro Thr Val Phe Ala Leu Leu Ser Lys 625 630 635 640 Asn Leu Met Ile ValHis Ser Asp Leu Ala Val His Phe Pro Ala Ile 645 650 655 Gln Tyr Ala ValLeu Tyr Thr Leu Tyr Ser His Cys Thr Arg His Asp 660 665 670 His Phe IleSer Ser Ser Leu Ser Ser Ala Ser Pro Ser Leu Phe Asp 675 680 685 Gly AlaVal Ile Ser Thr Val Thr Thr Ala Thr Lys Lys His Phe Ser 690 695 700 IleIle Leu Asn Leu Leu Gly Ile Leu Leu Lys Lys Asp Asn Leu Asn 705 710 715720 Gln Asp Thr Arg Lys Leu Leu Met Thr Trp Ala Leu Glu Ala Ala Val 725730 735 Leu Met Arg Lys Ser Glu Thr Tyr Ala Pro Leu Phe Ser Leu Pro Ser740 745 750 Phe His Lys Phe Cys Lys Gly Leu Leu Ala Asn Thr Leu Val GluAsp 755 760 765 Val Asn Ile Cys Leu Gln Ala Cys Ser Ser Leu His Ala LeuSer Ser 770 775 780 Ser Leu Pro Asp Asp Leu Leu Gln Arg Cys Val Asp ValCys Arg Val 785 790 795 800 Gln Leu Val His Ser Gly Thr Arg Ile Arg GlnAla Phe Gly Lys Leu 805 810 815 Leu Lys Ser Ile Pro Leu Asp Val Val LeuSer Asn Asn Asn His Thr 820 825 830 Glu Ile Gln Glu Ile Ser Leu Ala LeuArg Ser His Met Ser Lys Ala 835 840 845 Pro Ser Asn Thr Phe His Pro GlnAsp Phe Ser Asp Val Ile Ser Phe 850 855 860 Ile Leu Tyr Gly Asn Ser HisArg Thr Gly Lys Asp Asn Trp Leu Glu 865 870 875 880 Arg Leu Phe Tyr SerCys Gln Arg Leu Asp Lys Arg Asp Gln Ser Thr 885 890 895 Ile Pro Arg AsnLeu Leu Lys Thr Asp Ala Val Leu Trp Gln Trp Ala 900 905 910 Ile Trp GluAla Ala Gln Phe Thr Val Leu Ser Lys Leu Arg Thr Pro 915 920 925 Leu GlyArg Ala Gln Asp Thr Phe Gln Thr Ile Glu Gly Ile Ile Arg 930 935 940 SerLeu Ala Ala His Thr Leu Asn Pro Asp Gln Asp Val Ser Gln Trp 945 950 955960 Thr Thr Ala Asp Asn Asp Glu Gly His Gly Asn Asn Gln Leu Arg Leu 965970 975 Val Leu Leu Leu Gln Tyr Leu Glu Asn Leu Glu Lys Leu Met Tyr Asn980 985 990 Ala Tyr Glu Gly Cys Ala Asn Ala Leu Thr Ser Pro Pro Lys ValIle 995 1000 1005 Arg Thr Phe Phe Tyr Thr Asn Arg Gln Thr Cys Gln AspTrp Leu Thr 1010 1015 1020 Arg Ile Arg Leu Ser Ile Met Arg Val Gly LeuLeu Ala Gly Gln Pro 1025 1030 1035 1040 Ala Val Thr Val Arg His Gly PheAsp Leu Leu Thr Glu Met Lys Thr 1045 1050 1055 Thr Ser Leu Ser Gln GlyAsn Glu Leu Glu Val Thr Ile Met Met Val 1060 1065 1070 Val Glu Ala LeuCys Glu Leu His Cys Pro Glu Ala Ile Gln Gly Ile 1075 1080 1085 Ala ValTrp Ser Ser Ser Ile Val Gly Lys Asn Leu Leu Trp Ile Asn 1090 1095 1100Ser Val Ala Gln Gln Ala Glu Gly Arg Phe Glu Lys Ala Ser Val Glu 11051110 1115 1120 Tyr Gln Glu His Leu Cys Ala Met Thr Gly Val Asp Cys CysIle Ser 1125 1130 1135 Ser Phe Asp Lys Ser Val Leu Thr Leu Ala Asn AlaGly Arg Asn Ser 1140 1145 1150 Ala Ser Pro Lys His Ser Leu Asn Gly GluSer Arg Lys Thr Val Leu 1155 1160 1165 Ser Lys Pro Thr Asp Ser Ser ProGlu Val Ile Asn Tyr Leu Gly Asn 1170 1175 1180 Lys Ala Cys Glu Phe TyrIle Ser Ile Ala Asp Trp Ala Ala Val Gln 1185 1190 1195 1200 Glu Trp GlnAsn Ala Ile His Asp Leu Lys Lys Ser Thr Ser Ser Thr 1205 1210 1215 SerLeu Asn Leu Lys Ala Asp Phe Asn Tyr Ile Lys Ser Leu Ser Ser 1220 12251230 Phe Glu Ser Gly Lys Phe Val Glu Cys Thr Glu Gln Leu Glu Leu Leu1235 1240 1245 Pro Gly Glu Asn Ile Asn Leu Leu Ala Gly Gly Ser Lys GluLys Ile 1250 1255 1260 Asp Met Lys Lys Leu Leu Pro Asn Met Leu Ser ProAsp Pro Arg Glu 1265 1270 1275 1280 Leu Gln Lys Ser Ile Glu Val Gln LeuLeu Arg Ser Ser Val Cys Leu 1285 1290 1295 Ala Thr Ala Leu Asn Pro IleGlu Gln Asp Gln Lys Trp Gln Ser Ile 1300 1305 1310 Thr Glu Asn Val ValLys Tyr Leu Lys Gln Thr Ser Arg Ile Ala Ile 1315 1320 1325 Gly Pro LeuArg Leu Ser Thr Leu Thr Val Ser Gln Ser Leu Pro Val 1330 1335 1340 LeuSer Thr Leu Gln Leu Tyr Cys Ser Ser Ala Leu Glu Asn Thr Val 1345 13501355 1360 Ser Asn Arg Leu Ser Thr Glu Asp Cys Leu Ile Pro Leu Phe SerGlu 1365 1370 1375 Ala Leu Arg Ser Cys Lys Gln His Asp Val Arg Pro TrpMet Gln Ala 1380 1385 1390 Leu Arg Tyr Thr Met Tyr Gln Asn Gln Leu LeuGlu Lys Ile Lys Glu 1395 1400 1405 Gln Thr Val Pro Ile Arg Ser His LeuMet Glu Leu Gly Leu Thr Ala 1410 1415 1420 Ala Lys Phe Ala Arg Lys ArgGly Asn Val Ser Leu Ala Thr Arg Leu 1425 1430 1435 1440 Leu Ala Gln CysSer Glu Val Gln Leu Gly Lys Thr Thr Thr Ala Gln 1445 1450 1455 Asp LeuVal Gln His Phe Lys Lys Leu Ser Thr Gln Gly Gln Val Asp 1460 1465 1470Glu Lys Trp Gly Pro Glu Leu Asp Ile Glu Lys Thr Lys Leu Leu Tyr 14751480 1485 Thr Ala Gly Gln Ser Thr His Ala Met Glu Met Leu Ser Ser CysAla 1490 1495 1500 Ile Ser Phe Cys Lys Ser Val Lys Ala Glu Tyr Ala ValAla Lys Ser 1505 1510 1515 1520 Ile Leu Thr Leu Ala Lys Trp Ile Gln AlaGlu Trp Lys Glu Ile Ser 1525 1530 1535 Gly Gln Leu Lys Gln Val Tyr ArgAla Gln His Gln Gln Asn Phe Thr 1540 1545 1550 Gly Leu Ser Thr Leu SerLys Asn Ile Leu Thr Leu Ile Glu Leu Pro 1555 1560 1565 Ser Val Asn ThrMet Glu Glu Glu Tyr Pro Arg Ile Glu Ser Glu Ser 1570 1575 1580 Thr ValHis Ile Gly Val Gly Glu Pro Asp Phe Ile Leu Gly Gln Leu 1585 1590 15951600 Tyr His Leu Ser Ser Val Gln Ala Pro Glu Val Ala Lys Ser Trp Ala1605 1610 1615 Ala Leu Ala Ser Trp Ala Tyr Arg Trp Gly Arg Lys Val ValAsp Asn 1620 1625 1630 Ala Ser Gln Gly Glu Gly Val Arg Leu Leu Pro ArgGlu Lys Ser Glu 1635 1640 1645 Val Gln Asn Leu Leu Pro Asp Thr Ile ThrGlu Glu Glu Lys Glu Arg 1650 1655 1660 Ile Tyr Gly Ile Leu Gly Gln AlaVal Cys Arg Pro Ala Gly Ile Gln 1665 1670 1675 1680 Asp Glu Asp Ile ThrLeu Gln Ile Thr Glu Ser Glu Asp Asn Glu Glu 1685 1690 1695 Asp Asp MetVal Asp Val Ile Trp Arg Gln Leu Ile Ser Ser Cys Pro 1700 1705 1710 TrpLeu Ser Glu Leu Asp Glu Ser Ala Thr Glu Gly Val Ile Lys Val 1715 17201725 Trp Arg Lys Val Val Asp Arg Ile Phe Ser Leu Tyr Lys Leu Ser Cys1730 1735 1740 Ser Ala Tyr Phe Thr Phe Leu Lys Leu Asn Ala Gly Gln IlePro Leu 1745 1750 1755 1760 Asp Glu Asp Asp Pro Arg Leu His Leu Ser HisArg Val Glu Gln Ser 1765 1770 1775 Thr Asp Asp Met Ile Val Met Ala ThrLeu Arg Leu Leu Arg Leu Leu 1780 1785 1790 Val Lys His Ala Gly Glu LeuArg Gln Tyr Leu Glu His Gly Leu Glu 1795 1800 1805 Thr Thr Pro Thr AlaPro Trp Arg Gly Ile Ile Pro Gln Leu Phe Ser 1810 1815 1820 Arg Leu AsnHis Pro Glu Val Tyr Val Arg Gln Ser Ile Cys Asn Leu 1825 1830 1835 1840Leu Cys Arg Val Ala Gln Asp Ser Pro His Leu Ile Leu Tyr Pro Ala 18451850 1855 Ile Val Gly Thr Ile Ser Leu Ser Ser Glu Ser Gln Ala Ser GlyAsn 1860 1865 1870 Lys Phe Ser Thr Ala Ile Pro Thr Leu Leu Gly Asn IleGln Gly Glu 1875 1880 1885 Glu Leu Leu Val Ser Glu Cys Glu Gly Gly SerPro Pro Ala Ser Gln 1890 1895 1900 Asp Ser Asn Lys Asp Glu Pro Lys SerGly Leu Asn Glu Asp Gln Ala 1905 1910 1915 1920 Met Met Gln Asp Cys TyrSer Lys Ile Val Asp Lys Leu Ser Ser Ala 1925 1930 1935 Asn Pro Thr MetVal Leu Gln Val Gln Met Leu Val Ala Glu Leu Arg 1940 1945 1950 Arg ValThr Val Leu Trp Asp Glu Leu Trp Leu Gly Val Leu Leu Gln 1955 1960 1965Gln His Met Tyr Val Leu Arg Arg Ile Gln Gln Leu Glu Asp Glu Val 19701975 1980 Lys Arg Val Gln Asn Asn Asn Thr Leu Arg Lys Glu Glu Lys IleAla 1985 1990 1995 2000 Ile Met Arg Glu Arg His Thr Ala Leu Met Lys ProIle Val Phe Ala 2005 2010 2015 Leu Glu His Val Arg Ser Ile Thr Ala AlaPro Ala Glu Thr Pro His 2020 2025 2030 Glu Lys Trp Phe Gln Asp Asn TyrGly Asp Ala Ile Glu Asn Ala Leu 2035 2040 2045 Glu Lys Leu Lys Thr ProLeu Asn Pro Ala Lys Pro Gly Ser Ser Trp 2050 2055 2060 Ile Pro Phe LysGlu Ile Met Leu Ser Leu Gln Gln Arg Ala Gln Lys 2065 2070 2075 2080 ArgAla Ser Tyr Ile Leu Arg Leu Glu Glu Ile Ser Pro Trp Leu Ala 2085 20902095 Ala Met Thr Asn Thr Glu Ile Ala Leu Pro Gly Glu Val Ser Ala Arg2100 2105 2110 Asp Thr Val Thr Ile His Ser Val Gly Gly Thr Ile Thr IleLeu Pro 2115 2120 2125 Thr Lys Thr Lys Pro Lys Lys Leu Leu Phe Leu GlySer Asp Gly Lys 2130 2135 2140 Ser Tyr Pro Tyr Leu Phe Lys Gly Leu GluAsp Leu His Leu Asp Glu 2145 2150 2155 2160 Arg Ile Met Gln Phe Leu SerIle Val Asn Thr Met Phe Ala Thr Ile 2165 2170 2175 Asn Arg Gln Glu ThrPro Arg Phe His Ala Arg His Tyr Ser Val Thr 2180 2185 2190 Pro Leu GlyThr Arg Ser Gly Leu Ile Gln Trp Val Asp Gly Ala Thr 2195 2200 2205 ProLeu Phe Gly Leu Tyr Lys Arg Trp Gln Gln Arg Glu Ala Ala Leu 2210 22152220 Gln Ala Gln Lys Ala Gln Asp Ser Tyr Gln Thr Pro Gln Asn Pro Gly2225 2230 2235 2240 Ile Val Pro Arg Pro Ser Glu Leu Tyr Tyr Ser Lys IleGly Pro Ala 2245 2250 2255 Leu Lys Thr Val Gly Leu Ser Leu Asp Val SerArg Arg Asp Trp Pro 2260 2265 2270 Leu His Val Met Lys Ala Val Leu GluGlu Leu Met Glu Ala Thr Pro 2275 2280 2285 Pro Asn Leu Leu Ala Lys GluLeu Trp Ser Ser Cys Thr Thr Pro Asp 2290 2295 2300 Glu Trp Trp Arg ValThr Gln Ser Tyr Ala Arg Ser Thr Ala Val Met 2305 2310 2315 2320 Ser MetVal Gly Tyr Ile Ile Gly Leu Gly Asp Arg His Leu Asp Asn 2325 2330 2335Val Leu Ile Asp Met Thr Thr Gly Glu Val Val His Ile Asp Tyr Asn 23402345 2350 Val Cys Phe Glu Lys Gly Lys Ser Leu Arg Val Pro Glu Lys ValPro 2355 2360 2365 Phe Arg Met Thr Gln Asn Ile Glu Thr Ala Leu Gly ValThr Gly Val 2370 2375 2380 Glu Gly Val Phe Arg Leu Ser Cys Glu Gln ValLeu His Ile Met Arg 2385 2390 2395 2400 Arg Gly Arg Glu Thr Leu Leu ThrLeu Leu Glu Ala Phe Val Tyr Asp 2405 2410 2415 Pro Leu Val Asp Trp ThrAla Gly Gly Glu Ala Gly Phe Ala Gly Ala 2420 2425 2430 Val Tyr Gly GlyGly Gly Gln Gln Ala Glu Ser Lys Gln Ser Lys Arg 2435 2440 2445 Glu MetGlu Arg Glu Ile Thr Arg Ser Leu Phe Ser Ser Arg Val Ala 2450 2455 2460Glu Ile Lys Val Asn Trp Phe Lys Asn Arg Asp Glu Met Leu Val Val 24652470 2475 2480 Leu Pro Lys Leu Asp Gly Ser Leu Asp Glu Tyr Leu Ser LeuGln Glu 2485 2490 2495 Gln Leu Thr Asp Val Glu Lys Leu Gln Gly Lys LeuLeu Glu Glu Ile 2500 2505 2510 Glu Phe Leu Glu Gly Ala Glu Gly Val AspHis Pro Ser His Thr Leu 2515 2520 2525 Gln His Arg Tyr Ser Glu His ThrGln Leu Gln Thr Gln Gln Arg Ala 2530 2535 2540 Val Gln Glu Ala Ile GlnVal Lys Leu Asn Glu Phe Glu Gln Trp Ile 2545 2550 2555 2560 Thr His TyrGln Ala Ala Phe Asn Asn Leu Glu Ala Thr Gln Leu Ala 2565 2570 2575 SerLeu Leu Gln Glu Ile Ser Thr Gln Met Asp Leu Gly Pro Pro Ser 2580 25852590 Tyr Val Pro Ala Thr Ala Phe Leu Gln Asn Ala Gly Gln Ala His Leu2595 2600 2605 Ile Ser Gln Cys Glu Gln Leu Glu Gly Glu Val Gly Ala LeuLeu Gln 2610 2615 2620 Gln Arg Arg Ser Val Leu Arg Gly Cys Leu Glu GlnLeu His His Tyr 2625 2630 2635 2640 Ala Thr Val Ala Leu Gln Tyr Pro LysAla Ile Phe Gln Lys His Arg 2645 2650 2655 Ile Glu Gln Trp Lys Thr TrpMet Glu Glu Leu Ile Cys Asn Thr Thr 2660 2665 2670 Val Glu Arg Cys GlnGlu Leu Tyr Arg Lys Tyr Glu Met Gln Tyr Ala 2675 2680 2685 Pro Gln ProPro Pro Thr Val Cys Gln Phe Ile Thr Ala Thr Glu Met 2690 2695 2700 ThrLeu Gln Arg Tyr Ala Ala Asp Ile Asn Ser Arg Leu Ile Arg Gln 2705 27102715 2720 Val Glu Arg Leu Lys Gln Glu Ala Val Thr Val Pro Val Cys GluAsp 2725 2730 2735 Gln Leu Lys Glu Ile Glu Arg Cys Ile Lys Val Phe LeuHis Glu Asn 2740 2745 2750 Gly Glu Glu Gly Ser Leu Ser Leu Ala Ser ValIle Ile Ser Ala Leu 2755 2760 2765 Cys Thr Leu Thr Arg Arg Asn Leu MetMet Glu Gly Ala Ala Ser Ser 2770 2775 2780 Ala Gly Glu Gln Leu Val AspLeu Thr Ser Arg Asp Gly Ala Trp Phe 2785 2790 2795 2800 Leu Glu Glu LeuCys Ser Met Ser Gly Asn Val Thr Cys Leu Val Gln 2805 2810 2815 Leu LeuLys Gln Cys His Leu Val Pro Gln Asp Leu Asp Ile Pro Asn 2820 2825 2830Pro Met Glu Ala Ser Glu Thr Val His Leu Ala Asn Gly Val Tyr Thr 28352840 2845 Ser Leu Gln Glu Leu Asn Ser Asn Phe Arg Gln Ile Ile Phe ProGlu 2850 2855 2860 Ala Leu Arg Cys Leu Met Lys Gly Glu Tyr Thr Leu GluSer Met Leu 2865 2870 2875 2880 His Glu Leu Asp Gly Leu Ile Glu Gln ThrThr Asp Gly Val Pro Leu 2885 2890 2895 Gln Thr Leu Val Glu Ser Leu GlnAla Tyr Leu Arg Asn Ala Ala Met 2900 2905 2910 Gly Leu Glu Glu Glu ThrHis Ala His Tyr Ile Asp Val Ala Arg Leu 2915 2920 2925 Leu His Ala GlnTyr Gly Glu Leu Ile Gln Pro Arg Asn Gly Ser Val 2930 2935 2940 Asp GluThr Pro Lys Met Ser Ala Gly Gln Met Leu Leu Val Ala Phe 2945 2950 29552960 Asp Gly Met Phe Ala Gln Val Glu Thr Ala Phe Ser Leu Leu Val Glu2965 2970 2975 Lys Leu Asn Lys Met Glu Ile Pro Ile Ala Trp Arg Lys IleAsp Ile 2980 2985 2990 Ile Arg Glu Ala Arg Ser Thr Gln Val Asn Phe PheAsp Asp Asp Asn 2995 3000 3005 His Arg Gln Val Leu Glu Glu Ile Phe PheLeu Lys Arg Leu Gln Thr 3010 3015 3020 Ile Lys Glu Phe Phe Arg Leu CysGly Thr Phe Ser Lys Thr Leu Ser 3025 3030 3035 3040 Gly Ser Ser Ser LeuGlu Asp Gln Asn Thr Val Asn Gly Pro Val Gln 3045 3050 3055 Ile Val AsnVal Lys Thr Leu Phe Arg Asn Ser Cys Phe Ser Glu Asp 3060 3065 3070 GlnMet Ala Lys Pro Ile Lys Ala Phe Thr Ala Asp Phe Val Arg Gln 3075 30803085 Leu Leu Ile Gly Leu Pro Asn Gln Ala Leu Gly Leu Thr Leu Cys Ser3090 3095 3100 Phe Ile Ser Ala Leu Gly Val Asp Ile Ile Ala Gln Val GluAla Lys 3105 3110 3115 3120 Asp Phe Gly Ala Glu Ser Lys Val Ser Val AspAsp Leu Cys Lys Lys 3125 3130 3135 Ala Val Glu His Asn Ile Gln Ile GlyLys Phe Ser Gln Leu Val Met 3140 3145 3150 Asn Arg Ala Thr Val Leu AlaSer Ser Tyr Asp Thr Ala Trp Lys Lys 3155 3160 3165 His Asp Leu Val ArgArg Leu Glu Thr Ser Ile Ser Ser Cys Lys Thr 3170 3175 3180 Ser Leu GlnArg Val Gln Leu His Ile Ala Met Phe Gln Trp Gln His 3185 3190 3195 3200Glu Asp Leu Leu Ile Asn Arg Pro Gln Ala Met Ser Val Thr Pro Pro 32053210 3215 Pro Arg Ser Ala Ile Leu Thr Ser Met Lys Lys Lys Leu His ThrLeu 3220 3225 3230 Ser Gln Ile Glu Thr Ser Ile Ala Thr Val Gln Glu LysLeu Ala Ala 3235 3240 3245 Leu Glu Ser Ser Ile Glu Gln Arg Leu Lys TrpAla Gly Gly Ala Asn 3250 3255 3260 Pro Ala Leu Ala Pro Val Leu Gln AspPhe Glu Ala Thr Ile Ala Glu 3265 3270 3275 3280 Arg Arg Asn Leu Val LeuLys Glu Ser Gln Arg Ala Ser Gln Val Thr 3285 3290 3295 Phe Leu Cys SerAsn Ile Ile His Phe Glu Ser Leu Arg Thr Arg Thr 3300 3305 3310 Ala GluAla Leu Asn Leu Asp Ala Ala Leu Phe Glu Leu Ile Lys Arg 3315 3320 3325Cys Gln Gln Met Cys Ser Phe Ala Ser Gln Phe Asn Ser Ser Val Ser 33303335 3340 Glu Leu Glu Leu Arg Leu Leu Gln Arg Val Asp Thr Gly Leu GluHis 3345 3350 3355 3360 Pro Ile Gly Ser Ser Glu Trp Leu Leu Ser Ala HisLys Gln Leu Thr 3365 3370 3375 Gln Asp Met Ser Thr Gln Arg Ala Ile GlnThr Glu Lys Glu Gln Gln 3380 3385 3390 Ile Glu Thr Val Cys Glu Thr IleGln Asn Leu Val Asp Asn Ile Lys 3395 3400 3405 Thr Val Leu Thr Gly HisAsn Arg Gln Leu Gly Asp Val Lys His Leu 3410 3415 3420 Leu Lys Ala MetAla Lys Asp Glu Glu Ala Ala Leu Ala Asp Gly Glu 3425 3430 3435 3440 AspVal Pro Tyr Glu Asn Ser Val Arg Gln Phe Leu Gly Glu Tyr Lys 3445 34503455 Ser Trp Gln Asp Asn Ile Gln Thr Val Leu Phe Thr Leu Val Gln Ala3460 3465 3470 Met Gly Gln Val Arg Ser Gln Glu His Val Glu Met Leu GlnGlu Ile 3475 3480 3485 Thr Pro Thr Leu Lys Glu Leu Lys Thr Gln Ser GlnSer Ile Tyr Asn 3490 3495 3500 Asn Leu Val Ser Phe Ala Ser Pro Leu ValThr Asp Ala Thr Asn Glu 3505 3510 3515 3520 Cys Ser Ser Pro Thr Ser SerAla Thr Tyr Gln Pro Ser Phe Ala Ala 3525 3530 3535 Ala Val Arg Ser AsnThr Gly Gln Lys Thr Gln Pro Asp Val Met Ser 3540 3545 3550 Gln Asn AlaArg Lys Leu Ile Gln Lys Asn Leu Ala Thr Ser Ala Asp 3555 3560 3565 ThrPro Pro Ser Thr Val Pro Gly Thr Gly Lys Ser Val Ala Cys Ser 3570 35753580 Pro Lys Lys Ala Val Arg Asp Pro Lys Thr Gly Lys Ala Val Gln Glu3585 3590 3595 3600 Arg Asn Ser Tyr Ala Val Ser Val Trp Lys Arg Val LysAla Lys Leu 3605 3610 3615 Glu Gly Arg Asp Val Asp Pro Asn Arg Arg MetSer Val Ala Glu Gln 3620 3625 3630 Val Asp Tyr Val Ile Lys Glu Ala ThrAsn Leu Asp Asn Leu Ala Gln 3635 3640 3645 Leu Tyr Glu Gly Trp Thr AlaTrp Val 3650 3655 11 20 DNA Artificial Sequence syntheticoligonucleotide 11 agcaagctcc ctcctgtctc 20

What is claimed is:
 1. An isolated nucleic acid molecule, comprisingsubstantially the same nucleotide sequence as SEQ ID NO:1.
 2. Theisolated nucleic acid molecule of claim 1, wherein said nucleic acidmolecule encodes an ATX polypeptide comprising an amino acid sequenceshown in SEQ ID NO:2.
 3. A vector, comprising the isolated nucleic acidmolecule of claim
 1. 4. A host cell, comprising the vector of claim 3.5. A method of producing an ATX polypeptide comprising: a) growing thehost cell according to claim 4 under conditions appropriate forexpression of the ATX polypeptide, and b) isolating the ATX polypeptidefrom the host cell or host cell growth medium.
 6. An isolatedoligonucleotide, comprising at least 15 contiguous nucleotides of anucleotide sequence referenced as SEQ ID NO:11.
 7. An isolatedpolypeptide, comprising substantially the same amino acid sequence asSEQ ID NO:2.
 8. The polypeptide of claim 7, wherein said polypeptidecomprises an amino acid sequence as referenced in SEQ ID NO:2.
 9. Anantibody, or antigen binding fragment thereof, which specifically bindsto an ATX polypeptide comprising an amino acid sequence as referenced inSEQ ID NO:2.
 10. A method for identifying a compound that specificallybinds to an ATX polypeptide of claim 7, comprising: a) contacting saidATX polypeptide with a compound, and b) determining specific binding ofsaid compound to said ATX polypeptide.
 11. The method of claim 10,wherein said compound is a polypeptide.
 12. A method for identifying anATX-modulatory compound, comprising measuring the level of an ATXpolypeptide in the presence of a test compound, wherein a difference inthe level of said ATX polypeptide in the presence of said test compoundcompared to in the absence of said test compound indicating that saidtest compound is an ATX-modulatory compound, and wherein saidATX-modulatory compound is not caffeine or wortmannin.
 13. The method ofclaim 12, wherein said ATX-modulatory compound decreases the level ofATX polypeptide.
 14. The method of claim 12, wherein said ATX-modulatorycompound increases the level of ATX polypeptide.
 15. The method of claim12, wherein said level of ATX polypeptide is measured by determining thekinase activity of said ATX polypeptide.
 16. The method of claim 12,wherein said level of ATX polypeptide is measured by determining thephosphorylation of a p53 polypeptide or fragment.
 17. The method ofclaim 12, wherein said level of ATX polypeptide is measured bydetermining the level of p53 polypeptide accumulation.
 18. The method ofclaim 12, wherein said level of ATX polypeptide is measured bydetermining the level of non-sense mediated messenger RNA (mRNA) decay(NMD).
 19. The method of claim 12, wherein said ATX-modulatory compoundis an interfering RNA.
 20. A method for modulating cell survival,comprising introducing a compound identified by the method of claim 12into a cell in an amount effective to modulate survival of said cell.21. The method of claim 20, wherein said compound decreases cellsurvival.
 22. The method of claim 20, wherein said compound increasescell survival.
 23. The method of claim 20, wherein said cell is exposedto a stressor agent.
 24. The method of claim 23, wherein said stressoragent is selected from the group consisting of: UV light, ionizingradiation, and a chemical agent.
 25. A method for decreasing cellsurvival, comprising introducing the antisense oligonucleotide accordingto claim 6 into a cell in an amount effective to decrease survival ofsaid cell.